Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

Users Online : 191464

AbstractMaterial and MethodsResultsDiscussionReferencesDOI and Others
Article in PDF How to Cite Citation Manager Readers' Comments (0) Audio Visual Article Statistics Link to PUBMED Print this Article Send to a Friend
Advertisers Access Statistics Resources

Dr Mohan Z Mani

"Thank you very much for having published my article in record time.I would like to compliment you and your entire staff for your promptness, courtesy, and willingness to be customer friendly, which is quite unusual.I was given your reference by a colleague in pathology,and was able to directly phone your editorial office for clarifications.I would particularly like to thank the publication managers and the Assistant Editor who were following up my article. I would also like to thank you for adjusting the money I paid initially into payment for my modified article,and refunding the balance.
I wish all success to your journal and look forward to sending you any suitable similar article in future"



Dr Mohan Z Mani,
Professor & Head,
Department of Dermatolgy,
Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



Dr Kalyani R
Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
As an experienced dentist and an academician, I proudly recommend this journal to the dental fraternity as a good quality open access platform for rapid communication of their cutting-edge research progress and discovery.
I wish JCDR a great success and I hope that journal will soar higher with the passing time."



Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




Dr. Arunava Biswas

"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."



Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2012 | Month : May | Volume : 6 | Issue : 4 | Page : 636 - 641 Full Version

Comparative Evaluation of the Diagnostic Tests for Helicobacter pylori and Dietary Influence for Its Acquisition in Dyspeptic Patients: A Rural Hospital Based Study in Central India


Published: May 1, 2012 | DOI: https://doi.org/10.7860/JCDR/2012/.2152
Navinchandra Motiram Kaore, Neena Vinay Nagdeo, Vilas R. Thombare

1. Assistant Professor, Department of Microbiology, Peoples College of Medical sciences & RC, Bhopal India-462037. 2. Associate Professor, Department of Microbiology, NKP Salve Institute of Medical Sciences & RC, Nagpur, India. 3. Professor & Head of Department, Microbiology, NKP Salve Institute of Medical Sciences & RC, Nagpur, India.

Correspondence Address :
Dr. Navinchandra M. Kaore
HIG A-13, Peoples College of Medical Sciences & RC,
Bhanpur Road, Bhopal, India - 462037.
Madhya Pradesh, India.
Phone: +91-9584354161
E-mail: navinnmk@gmail.com; navinnmk@rediffmail.com

Abstract

Introduction: Dyspepsia is a very common group of symptoms referring upper gastrointestinal tract for which patients consult the physician, accounting for about 4% to 14% of their consultations. Annual Incidence of Helicobacter pylori is 6%-14% in developing countries with faeco-oral route of transmission. In this context this study has been undertaken to look for the association of Helicobacter pylori in patients of dyspepsia with comparative evaluation of diagnostic test for detection of helicobacter pylori and also look for effects of smoking and alcohol consumption over symptomatic colonization in dyspeptic patients attending the NKP Salve Institute of Medical Sciences and Research Centre, Digdoh hills, Nagpur, a tertiary care hospital catering to rural population of central India.

Materials & Methods: It was a cross sectional prospective study involving 92 patients. After obtaining history of their dietary and personal habits, 3 mucosal biopsy specimens were collected from pyloric end of the stomach along with blood sample. Specimens were processed immediately for culture, direct gram staining, rapid urease test, histology and serology along with Urea breath test by HeliprobeTM.

All the data was managed in Microsoft Excel and statistical analysis was done using Epi-info. Pearson’s chi square test was used for comparative evaluation between two groups.

Results: 55% patients were considered as positive by the three test criteria and taken as Reference Standard as being infected by H. pylori. Diagnostic tests used as well as dietary habits were evaluated against the reference standard.

Conclusion: Culture and in absence of culture; RUT, Direct Gram Stain, and Serology in combination, followed by RUT, Direct Gram Staining with Histology are the best for increasing the specificity for diagnosis of Helicobacter pylori infection in patients of dyspepsia. Alcohol consumption is not significantly associated but smoking and tobacco chewing or smoking along with alcohol consumption is highly significant for symptomatic colonization of H. pylori in dyspeptic patients.Key-words: Helicobacter pylori, Dyspepsia, Reference Standard, Evaluation of diagnostic tests, Dietary influence, Smoking, Alcohol.

Keywords

Helicobacter pylori, Dyspepsia, Reference Standard, Evaluation of diagnostic tests, Dietary influence, Smoking, Alcohol

Introduction
Dyspepsia is a very common human experience for which there are numerous causes. Dyspepsia is the group of symptoms which refers to the upper gastrointestinal tract for which patients consult the physician, accounting for about 4% to 14% of their consultations. However, the prevalence of dyspepsia in the general population is much higher, with 20%-30% patients experiencing dyspeptic symptoms and with many of them self medicating themselves (1),(2).

Until the discovery of Helicobacter pylori, the physicians and microbiologists believed that the stomach was likely to be sterile because of the presence of its acid ‘milieu’. The successful isolation of spiral gram negative bacilli by them has changed the focus from a non-infectious to an infectious aetiology i.e. from pH to Hp (3).

H. pylori is common bacterium and approximately 50 % population of the world is said to be infected (4). Humans are the only known reservoirs of H. pylori and the risk factors which have been described for acquiring the infection include residence in a developing country, poor socioeconomic conditions, overcrowding and an ethnic and genetic predisposition. Upto 80% of the children under the age of 10 years are infected in the developing countries. The prevalence of this infection in India is 22%, 56% and 87% in the 0-4 years, 5-9 years and in the 10-19 years age group respectively (5).

The annual incidence of the Helicobacter pylori infection is 0.3% to 0.7 % in the developed countries and it is 6-14 % in the developing countries (6).

The accepted routes of its transmission are the faeco-oral route in the developing countries and the gastro-oral route (through the vomitus and the saliva) in the developed countries (7).

Today, the role of Helicobacter pylori has been established in chronic antral gastritis, duodenal ulcers, chronic gastric ulcers, dyspepsia, gastric cancer and gastric lymphoma. The World Health Organization has added H. pylori to its list of known carcinogens (8).

There is a paucity of data on the incidence of Helicobacter pylori in patients of dyspepsia from the rural areas of central India. The literature about the dietary influence of alcohol consumption, smoking and in particular, tobacco chewing, which is not common in the western world, is also lacking. These habits are commonly prevalent in the rural areas and it is necessary to study their actual role in symptomatic colonization.

In this context, this study was undertaken to look for the association of Helicobacter pylori in patients of dyspepsia, with a comparative evaluation of the diagnostic tests for the detection of Helicobacter pylori and also to look for the dietary influence of alcohol consumption, smoking and tobacco chewing over the symptomatic colonization in dyspeptic patients who attended the tertiary care hospital which catered to the rural population of central India.

Material and Methods

This cross sectional, prospective study was carried out in the Department of Microbiology from December 2006 to November 2008 after obtaining permission from the institutional ethical committee. Patients of either sex and of any age who had the symptoms of dyspepsia were selected for the study. After obtaining the informed written consent, the patient’s details were noted down according to the case record form, including their dietary and personal habits. The chronic consumption of alcohol and tobacco along with their smoking habits were identified.

Collection of the Specimen
The diagnostic modalities were considered under:
(1) Invasive
  (a) Culture
  (b) Microscopic examination:
     (i) Direct Gram Staining
    (ii) Histopathology
  (c) Biopsy urease or Rapid Urease Test
(2) Non-invasive
  (a) Urea Breath Test
  (b) Serology

The patients were taken for upper G.I. Endoscopy after keeping them nil by mouth overnight. The patients who were on proton pump inhibitors were asked to discontinue the same for 10 days before being given an appointment for upper G.I. endoscopy, as it decreased the diagnostic yield.

Collection and Transport of the Biopsy specimen
Three mucosal biopsy specimens were collected from the pyloric (antral) end of the stomach.

Two specimens were taken in 0.2 ml of normal physiological saline which was just enough to keep them moist for the culture, direct gram staining and for the rapid urease test (RUT). The biopsies were transported within two hrs to the Microbiology Laboratory and in case of a delay, they were stored at 40C for 5-6 hours (9),(10).

One specimen was taken in 10% formalin for histopathology.

Collection of blood for serum
The blood samples were collected by taking all the aseptic precautions and they were transferred to a collecting tube till the serum separated. The separated serum was divided into small aliquots and these were properly labeled and stored at -20o C to be processed collectively for IgG against H. pylori later.

Processing of the specimens
All the samples were processed immediately after (within 2 hours) their transportation to the Microbiology Laboratory.

Culture
One biopsy specimen was homogenized by using a sterile mortar and pestle, in little sterile broth. This was used for the inoculation of the plates and for preparing the smears for gram staining.

The Brucella agar base and the Columbia agar base were procured from Himedia laboratories. The media were prepared by the addition of 5%-7% defibrinated sheep blood and they were made selective by the addition of the Campylobacter supplement (Skirrow’s) – FD- 008 from Himedia.

All the selective and the non-selective media were inoculated with homogenized material and they were incubated at 37°C in a candle jar with a pad of cotton which was soaked in water, which was placed at the bottom, to provide a humidified microaerophilic environment. The plates would be examined on the 3rd, 5th and the 7th day of their incubation (9),(11). Characteristic, shiny translucent colonies were identified by gram staining and the urease, catalase and the oxidase tests (12).

Direct Gram Staining
The smears were prepared on clean, grease free slides from the tissue homogenate which was fixed with 95 % ethanol, by putting 2-3 drops of it on the slide and allowing them to evaporate.

The smears which were thus prepared were stained by using Jensen’s modification of gram staining, but instead of using Saffranin, dilute Carbol fuschin (0.2 %) was used as a counterstain, which helped in the better visualization of gram negative, spiral or sea gull wing shaped bacteria, which were present mucosally and submucosally (10),(11),(13),(14).

Histopathology
The biopsy specimens which were collected in 10 % formalin as a fixative were transported to the Pathology Laboratory after proper labeling and with the requisition form for the Haematoxalin and eosin staining. After this, they were observed and reported by the pathologists (15).

The Rapid Urease test (RUT)
The Rapid urease or the Biopsy urease test was performed for putting the second biopsy specimen into 0.5 ml of freshly prepared unbuffered 10 % urea solution in de ionized water, with 1% phenol red as an indicator, whose pH was adjusted from 6.8 to 7.2. An uninoculated tube was also kept as a negative control. The inoculated and the control tubes were incubated and the results were read at the end of 1 hour, 4 hours and 24 hours. The hydrolysis of the urea liberated ammonia, which increased the pH and changed the colour of the indicator medium from yellow to pink, thus indicating a positive test (16),(17).

IgG against Helicobacter pylori
The EIAgen Helicobacter pylori IgG Kit from Adaltis Italia S.p.A., (Bologna) Italy-12, which was an ELIZA based kit, was used. All the serum samples which were stored at -20oC were thawed and all the test procedures were carried out according to the manufacturer’s instructions. Titres above the cut off value of 15 AU/ml were noted as positive.

The Urea Breath test
“Heliprobeâ€TM, which was marketed by Rad- Probe and developed by Kibion Diagnostic, Uppsala, Sweden, was used for the urea breath test.

The Heliprobe Procedure (18)
After a 6 hour fasting period, a 37- kBq14 C Urea capsule was given to the patients, with 50 ml of water. After 10 min, the breath samples are collected by making the patients blow into the Heliprobe Breath card till it got saturated, which was indicated by the change of colour of the indicator from orange to yellow. The change of colour indicated complete saturation of the CO2 in the LiOH (Lithium hydroxide) pads. These Heliprobe Breath Cards were the put into the slot of the Heliprobe Breath Analyzer. The analysis was based on the number of beta particles that hit the two Geiger Muller Counters during the 250 second measurement cycle and the results were presented as counts per minute (cpm), together with the test results- negative, equivocal or positive and they were reported as:

Heliprobe O : Negative (<25 cpm)
Heliprobe 1 : Equivocal (25 to 50 cpm)
Heliprobe 2 : Positive ( >50 cpm )

All the data was managed in Microsoft Excel and the statistical analysis was done by using Epi-info. The Pearson’s Chi square test was used for the comparative evaluation between the two groups.

Results

A total of ninety two antral (92) biopsy samples were obtained and 57 blood samples were collected for serum.

A male predominance of 58/92 (63.04%) was observed, as compared to that of 34/92 (36.96 %) in females. Most of the patients presented in the active age group of 20 to 40 years.

(Table/Fig 1) shows the number of samples which were processed by various diagnostic methods for the diagnosis of Helicobacter pylori and their overall sensitivities.

There is no failsafe method for the diagnosis of Helicobacter pylori with a sensitivity and specificity of 100 %. To increase the specificity, the three test criteria has been adopted for the diagnosis of the Helicobacter pylori infection. By this criteria, all the culture positive cases; and in the absence of culture positivity, patients with any three (3) diagnostic tests which are positive out of five (5) test that have been carried out were taken as being infected for Helicobacter pylori. A total of 55 patients were found to be positive either by culture or by any other three tests (Table/Fig 2).

These 55 patients were considered as positive (infected by H.pylori) by the three test criteria and they were taken as the Reference Standard.. The comparative evaluation of the different tests which were done to study the incidence of Helicobacter pylori in the patients of dyspepsia has been shown in (Table/Fig 3).

The effect of alcohol and tobacco chewing was also seen over the symptomatic colonization of Helicobacter pylori in dyspepsia. (Table/Fig 4) .

Discussion

In our study, a clear male predominance of 58/92 (63.04%) was observed, as compared to that of 34/92 (36.96 %) in females who presented with symptoms. The biological differences, whatever they are, must stem ultimately from the greater quantity of genetic material which is possessed by the female. The presence of one X chromosome in the male infant thus confers less immunological protection as compared to that in the female counterparts (19).

Similar findings were found in a study which was done in India by Ahmed et al., who also observed the active age group of 20 to 40 yrs to be more vulnerable, as was observed in our study (20).

8/92 samples were positive by culture with a sensitivity of 8.69%. A sensitivity of over 90% was reported by various workers like Marshall et al., Jones et al., and Goodwin et al., (9),(15),(21). In contrast, the Indian studies reported a sensitivity which ranged from 1.09% (as reported by A. Ayyagari) to 63% (as reported by Akbar and Eltahawy) (11), (22).

Our culture sensitivity was in accordance with these results. The low rate of isolation may be because of the fastidious nature of H. pylori and because of a number of other factors like the patchy distribution of the organism, inadequate mincing of the biopsy material, the presence of oropharyngeal flora, the loss of viability of the specimen during transportation, etc. These factors are difficult to control. All these factors together, result in low sensitivity and a low negative predictive value.

The rampant use of over the counter prescriptions of the antisecretory agents, as well as of Metronedazole for the protozoan infestations, which are quite common in the rural regions of India, decrease the positive outcome (23), (24).

Therefore, though it was considered as a gold standard, it is now used only in research settings (25).

76/92 (82.60%) samples in our study were positive by the Rapid urease test, which is in agreement with the findings of almost all the studies in which the sensitivity ranged from 72 % (as reported by U. arora et al) to 99 % (as reported by Pinit Kullavanijaya) (26).

With the use of un-buffered 10 % urea solution, our results were found to be in agreement with those of other workers like A.S.Arvind et al., U. arora et al., and Vaira D., et al (16),(17),(27).

A total of 65 samples were processed for the histo-pathoplogical examination by haematoxilin and eosin staining, out of which a total of 56 (86.15%) showed the changes of antral gastritis. Like in our study, most studies which have been reported in the western literature, which include those by Warren et al,, Goodwin et al., and Morris et al., have found H.pylori to be associated with chronic active gastritis (21),(28),(29).

A total of 92 biopsy specimens were processed by using modified gram staining in which the slides were counterstained by using dilute Carbol Fuschin. 72 out of the 92 samples (78.26%) were positive for Helicobacter pylori (Table/Fig 1).

Ayyagari reported the staining of the crushed tissue smears by Gram’s method by using dilute Carbol Fuschin as counter stain as the simplest and reliable technique with 80.8 % sensitivity. K.N. Prasad et al., reported a sensitivity of 34 % for the same method (11),(30).

D’Costa F Grace et al., found gram staining to be 74.2% sensitive. With 78.26% positivity, our findings were in accordance with those of other workers (31).

40/57 samples were positive (70.17%) for IgG against Helicobacter pylori by this ELISA based test. Pinit Kullavanijaya et al reported 120/191 patients to be positive i.e. 62.8 %, with a sensitivity of 96.8% and a specificity of 96.8 %, while Abida Malik et al reported a seropositivity of 58.3 % [26,23]. Daad H Akbar et al reported 212/289 (73%) patients to be positive for IgG (22). Our positivity rate of 70.17 % was comparable to that which was reported by all the authors.

For the Urea Breath test, the HELIPROBE system was procured late in the course of the study. Though it was subsidized, the higher cost was a constraint for the patients, as a rural population mainly attended our tertiary care unit. Thus, only 10 patients could be included for Heliprobe. 6/10 (60%) patients were found to be positive for the Urea Breath test.

The low rate of positivity in our study may be attributed to the low number of samples, over the counter prescription of the proton pump inhibitors and also to the use of Metronedazole for protozoan infestations, which were very common in the rural settings in India, which could increase the false negative cases (23), (24).

In the absence of a failsafe method, a battery of tests are required for increasing the specificity of the diagnosis (32). Many workers, in the past, have considered culture as a gold standard and in the absence of culture, a positive outcome of more than one test (invasive or non invasive) was taken as a reference standard for the diagnosis of the Helicobacter pylori infection. This kind of reference standard has been used by various workers (11),(22),(26),(29),(33),(34),(35).

A total of 55 patients were positive by the reference standard which has been described above. 40/58 (68.96%) males and 15/34 (44.11%) females in our study were found to be infected with Helicobacter pylori by using the three test criteria (Table/Fig 2).

There were 8 patients which were positive by culture. All the eight patients were also positive by the Rapid Urease Test (RUT), histology, direct gram staining and serology (22),(26).

55 out of 92 patients i.e. 58.78 % were diagnosed by the three test criteria (Reference standard). This rate was similar to that which was reported by other investigators like Bulk et al., who reported a rate of 69% and M.A. El Barrawy who reported a rate of 61% (35).

RUT and direct gram staining with the non invasive test of serology was more effective in combination in diagnosing 17/55 patients (30.90%), followed by RUT and direct gram staining with histology 16/55 (29.09%), as was reported also by Pinit Kullavanijaya et al., and Daad H. Akbar et al, followed by a combination of direct gram staining, serology and histology [8/55 (14.54%)] and culture [8/55 (14.54%)] (26),(22).

(Table/Fig 3) shows that culture and the Rapid Urease Test were significantly associated and that direct gram staining and IgG detection were highly significant for the diagnosis of H.pylori. (22),(26),(33),(34),(35).

Histology alone was not significant in the diagnosis of the Helicobacter pylori infection, but along with the combination of RUT and direct gram staining, histopathology was quite useful for its diagnosis (Table/Fig 2). The lower sensitivity and the lack of a significant association may be due to the small no of samples which were processed for histopathology as well as drug induced gastritis, because of the rampant use of NSAIDs which are easily available as over the counter prescriptions in rural India.

Good sensitivity and specificity were reported by the workers for Heliprobe. This is a new system and the time interval after which the patient exhales into the Heliprobe card also has an effect over the test positivity. Ideally, it should be 10 min and a delay may cause the 14C labeled urea from getting hydrolyzed by other urease producing organisms like Campylobacter jejuni. The protease which is present in the intestine can give false positive results (18), (36), (37).

The effect of alcohol, smoking and tobacco chewing in patients as a possible risk factor, was also evaluated for the symptomatic colonization of Helicobacter pylori in patients of dyspepsia by using the patients who were diagnosed by using a reference standard (Table/Fig 4).

With 16/30 patients being diagnosed by using the reference standard, the alcoholic patients were not significantly associated with the symptomatic colonization, while with 30/40 patients who were diagnosed as positive for the Helicobacter pylori infection had smoking or tobacco chewing habits. They were significantly associated with the symptomatic colonization, as was also reported by Cardenas VM et al., and el-Gueid A et al., (38),(39).

14/15 patients with chronic alcoholism along with tobacco chewing were found to be significantly associated with the symptomatic colonization. The increased C-X-C chemokine mRNA expression which was seen in smokers and tobacco chewers could play a role in inducing an enhanced inflammatory activity in gastritis and in the consequent severe disease which was associated with the Helicobacter pylori infection. Shimoyana T et al., and MA El- Barrawy reported similar findings (35),(40).

To conclude: In culture and in the absence of culture RUT, direct gram staining, and serology in combination was the best [17/55 patients (30.90%)], followed by RUT and direct gram staining with histology [16/55 (29.09%)] for increasing the specificity for the diagnosis of the Helicobacter pylori infection in patients of dyspepsia.

Alcohol consumption was not significantly associated with the symptomatic colonization of H.pylori and it was found to have a protective effect against the colonization.

Smoking and tobacco chewing or smoking along with alcohol consumption was highly significant for the symptomatic colonization of H. pylori in dyspeptic patients.

References

1.
Chiba N. Definition of dyspepsia: the time for reprisal. Eur J Surg supplement 1998; 583:14-23.
2.
Knill-Jones RP. A formal approach to the symptoms in dyspepsia. Clinical Gastroenterol 1985;14:517-29.
3.
Marshall BJ, Warren JR. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1984; i:1311-15.
4.
Brown LM. Helicobacter pylori: epidemiology and the routes of transmission. Epidemiol Rev 2000; 22(2): 283–97.
5.
Das JC, Paul N. The epidemiology and the pathophysiology of the Helicobacter pylori infection in children. Indian J Paediatr 2007;74: 287-90.
6.
Logan RPH, Walker MM. ABC of the upper gastrointestinal tract: the epidemiology and the diagnosis of the Helicobacter pylori infection. BMJ 2001; 323(7318): 920–22.
7.
Philippe L, Olzem Y. Epidemiology of the Helicobacter pylori infection: A year in Helicobacter 2007, Journal compilation, Blackwell Publishing Ltd, Helicobacter 12 (suppl. 1):1-3.
8.
Calam J. A clinician’s guide to Helicobacter pylori: Ist ed. (Chapman and Hall, London) 1996;72-78.
9.
Goodwin CS, Blincow E, Warren JR, Waters TE, Sanderson CR, Easton CR. Evaluation of cultural techniques for isolating Campylobacter pyloridis from endoscopic biopsies of the gastric mucosa. J Clin Pathol 1985;38:1127-31.
10.
Balkrissh Nair G, Bhattacharya SK, Dutta P, Sen JK, Dutta D, et al. Campylobacter pyloridis in samples of endoscopic biopsy in patients with upper gastrointestinal tract disease. Indian J Med Res 1986;84:574-76.
11.
Ayyagiri A, Ray P, Kochhar R, Bhasin D, Siddeshi ER, Singh K, et al. Evaluation of different methods for the detection of Helicobacter pylori in patients with gastric disease. Indian J Med Res. 1990;91:126-28.
12.
McNulty CAM, Dent JC. Rapid identification of Campylobacter pylori by using preformed enzymes. J Clin Microbiol 1987;25(9):1683-86.
13.
McNulty CAM, Dent JC, Uff JS, Gear MWL, Wilkinson SP. Detection of Campylobacter pylori by the biopsy urease test: an assessment in 1445 patients. Gut 1989;30:1058-62.
14.
Trowel JE, Yoong AKH, Sanl KJ, Gant PW, Bell GD. Simple half gram stain for showing the presence of Camplobacter pyloridis in sections. J Clin Pathol 1987;40:702.
15.
Jones DM, Lesselle AM, Eldridge J. Campylobacter like organisms on the gastric mucosa: culture, histological and serological studies. Schweiz Med Wochenschr 1988;118(9):293-301.
16.
Arvind AS, Cook RS, Tabaqchali S, Farthing MJG. A one minute endoscopy room test for Campylobacter pylori. Lancet 1988; i:704.
17.
Arora U, Aggarwal A, Singh K. A comparative evaluation of the conventional methods and the detection of the ELISA based IgG antibodies for the diagnosis of the Helicobacter pylori infection in cases of dyspepsia. Indian J Med Microbi 2003;21: 46-48.
18.
Ozturk E, Yesilova Z, Iigan S, Arslan N, Erdit A, Celasun B, et al. A new practical low dose 14C urea breath test for the diagnosis of the Helicobacter pylori infection : Clinical validation and comparison with the standard methods. Eur J Nucl Mol Imaging 2003; 30: 1457-62.
19.
Washburn T C, Medearis Jr. DN , Childs B . The sex differences in the susceptibility to infections. Paediatrics 1965;35(1): 57-64.
20.
Ahmed KS, Khan AA, Ahmed I, et al. A prevalence study to elucidate the transmission pathways of Helicobacter pylori at the oral and gastroduodenal sites of a south Indian population. Singapore Med J 2006;47:291-96.
21.
Marshall BJ, Royce H, Annear DI, Goodwin CS, Pearman JW, Warren JR, et al. Original isolation of Campylobacter pyloridis from human gastric mucosa. Microbios Lett. 1984;25:83-88.
22.
Akbar DH, Eltahawy ATA. Helicobacter pylori infection at a university hospital in Saudi Arabia :Prevalence and comparison of the diagnostic modalities and the endoscopic findings. Indian J Pathol Micr 2005;48(2):181-85.
23.
Malik A, Singhal S, Mukharji U. Evaluation of an ELISA based antibody test against H.pylori in cases of non-ulcer dyspepsia. J Med Microbiol 1999;17(3):137-39.
24.
American Gastroenterological Association. Technical review on the evaluation of dyspepsia. Gastroenterology 2005;129:1756-80.
25.
Alikhani MY, Sadeghifard N, Farajnia S, Hajja M, Aslani MM, Zamani AR. Evaluation of selective and non selective media for the isolation of Helicobacter pylori from gastric biopsy specimens. Pak J Bio Sci 2007;10(22):4156-59.
26.
Kullavanijaya P, Thong-Ngam D, Hanvivatvong O, Nunthapisud P, Tangkijvanich P, Suwanagool P. Analysis of eight different methods for the detection of the Helicobacter pylori infection in patients with dyspepsia. J Gastroenterol Hepatol 2004;19(12):1392-96.
27.
Vaira D, Holton J, Crirns S, Falzon M, Salmon P. The four hour rapid urease test (RUT) for detecting Campylobacter pylori: is it reliable enough to start the treatment?. J. Clin Pathol 1988;41:355-56.
28.
Goodwin CS, Armstrong JA, Marshall BJ. Campylobacter pyloridis, gastritis and peptic ulceration. J Clin Pathol 1986;39:353-65.
29.
Morris A, Nicholson G, Lloyd G, Haines D, Rogers A, Taylor D. Seroepidemiology of Campylobacter pyloridis. N Z Med J 1986;99:657–59.
30.
Prasad KN, Singhal S, Ayyagari A, Sharma S, Dhole TN. Association of Helicobacter pylori in patients with non ulcer dyspepsia. Indian J Microbiol 1991;31(4):415-18.
31.
D’costa FG, Datta S, Patil Y. Histopathology of gastro-duodenal biopsies and the Helicobacter pylori infection. Bombay Hospital Journal 2007; 292-300.
32.
Sengupta S, Saraswathi K, Varaiya A, De A, Gogate A. Helicobacter pylori in duodenal ulcer disease and its eradication. Indian J Med Microbiol 2002;20(3):163-64.
33.
Branka AM, De Wouw V, De Boer WA, Jansz A R, Roymans RTJM, Stalls APG. Comparison of three commercially available enzyme linked immunosorbent assays and a biopsy dependant diagnosis for detecting the Helicobacter pylori infection. J Clin Microbiol 1996; 94-97.
34.
Garcia-Diaz E, Castro-Fernandez M, Romero-Gomez M, Vargas- Romero J. The effectiveness of the IgG ELISA serology as an alternative diagnostic method for detecting the Helicobacter pylori infection in patients with gastro intestinal bleeding due to gastro-duodenal ulcers. Rev Esp Enferm Dig. 2002;94(12):725-36.
35.
El-Barrawy MA, Morod MI, Gaber M. The role of Helicobacter pylori in the genesis of gastric ulceration among smokers and non smokers. Eastern Mediterranean Health Journal 1997;2:316-21.
36.
Munster DJ, Chapman BA, Burt MJ, Dobbs BR, Allardyce RA, Bagshaw PF, et al. The fate of ingested 14C-urea in the urea breath test for the Helicobacter pylori infection. Scand J Gastroenterol 1993;28(8):661-66.
37.
Van Rauws EM, Tytgat GNT, van Royen EA. Cl4- The urea breath test for the non-invasive detection of the Campylobacter pylori colonization. Gastroentenology 1988;94:370-72.
38.
Cardenas VM, Graham DY. Smoking and the Helicobacter pylori infection in a sample of U.S. adults. Epidemiology 2005;16(4):586-90.
39.
el-guneid A,el-Sherif AM, Murray-Lyon IM, Zureikat N, Shousa S. The effect of chewing Qat on the mucosal histology and on the prevalence of Helicobacter pylori in the oesophagus, stomach and the duodenum of Yemeni patients. Hstopathology 1991;19(5):437-43.
40.
Shimoyama T, Everett SM, Fukuda S, Axon ATR, Dixon MF, Crabtree JE. Influence of smoking and alcohol on gastric chemokine mRNA expression in patients with Helicobacter pylori infection. J Clin Pathol 2001;54:332-34.

DOI and Others

ID: JCDR/2012/3727:0023

Date of Submission: Nov 28, 2011
Date of peer review: Jan 27, 2012
Date of acceptance: Feb 09, 2012
Date of Publishing: May 31, 2012

JCDR is now Monthly and more widely Indexed .
  • Emerging Sources Citation Index (Web of Science, thomsonreuters)
  • Index Copernicus ICV 2017: 134.54
  • Academic Search Complete Database
  • Directory of Open Access Journals (DOAJ)
  • Embase
  • EBSCOhost
  • Google Scholar
  • HINARI Access to Research in Health Programme
  • Indian Science Abstracts (ISA)
  • Journal seek Database
  • Google
  • Popline (reproductive health literature)
  • www.omnimedicalsearch.com