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

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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 : 2022 | Month : May | Volume : 16 | Issue : 5 | Page : DC01 - DC05 Full Version

A One Man Army- TrueNat Testing for the Identification of COVID-19 in Firozabad, Uttar Pradesh, India


Published: May 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/55742.16292
Lekha Tuli, Rohit Patawa

1. Associate Professor, Department of Microbiology, Autonomous State Medical College, Firozabad, Uttar Pradesh, India. 2. Guest Faculty, Department of Statistics, University of Allahabad, Allahabad, Uttar Pradesh, India.

Correspondence Address :
Dr. Lekha Tuli,
117/259,Q block,Sharda Nagar, Kanpur-208025, Uttar Pradesh, India.
E-mail: tuli_lekha@rediffmail.com

Abstract

Introduction: The Coronavirus Disease 2019 (COVID-19) pandemic has affected the entire world. The need of timely detection of the virus has been of prime importance and the efforts to develop sensitive, specific, rapid, portable and cost effective diagnostic methods promoted the indigenous development of TrueNat testing for viral load in COVID-19 detection which had been previously designed for detection of Tuberculosis and other infectious organisms.

Aim: To see the importance of TrueNat testing among symptomatic and asymptomatic cases in different age groups and gender.

Materials and Methods: In this retrospective study conducted in the Department of Microbiology, Autonomous State Medical College and SNM Hospital, Firozabad, Uttar Pradesh, India, from June 2020 - May 2021, a total of 4,659 samples were collected from patients (Influenza Like Illness (ILI), Severe Acute Respiratory Illness (SARI), symptomatic, asymptomatic, those seeking hospitalisation, emergency), contacts and travellers and were subjected to testing by TrueNat (Molbio Quattro). The cases were divided into group A of patients who presented with symptoms ≤7 days; group B of patients who presented with signs and symptoms >7 days and group C comprised of asymptomatic patients. The symptoms of patients were associated with the Cycle threshold (Ct) values of the Envelope (E) gene and the RNAdependent RNA polymerase gene (RdRp) gene. The Chi-square test was done to test the statistical significance of association of symptomatic patients with the outcome of the test.

Results: The maximum number of positive cases were found in the people 20-39 years (p-value <0.05). The least positivity was found in the higher (80 years) and lower (below 9 years) age groups. The positivity rates had no significant impact on the gender. The percentage positivity as detected by TrueNat testing was 3.3% and maximum positive patients were found in the group having symptoms <7 days (p<0.05). On association of the Ct values of E gene and RdRp gene with the symptoms it was found that 28.1% and 27.2% of the patients were in the high Ct value group.

Conclusion: TrueNat was found to be a portable and easy to perform test which did not require special laboratory set up. The use of Viral Lysis Medium (VLM) reduced the time of RNA extraction which not only rendered it safer to perform but expedited the results.

Keywords

Cartridge, Ct value, Molbio, Point of care

The beginning of the year 2020 has brought many unsolicited events for mankind. The COVID-19 pandemic has hit our lives like a tornado leaving many people helpless, devastated and stranded alone. The route of transmission of the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) virus is air borne, which makes its spread very easy thus affecting masses. Moreover, the clinical symptoms mimic those of flu or other respiratory infections and thus necessitate a prompt and accurate diagnosis. In view of this, the World Health Organisation (WHO) endorsed the rapid molecular TrueNat assay for detection of COVID-19 virus and Indian Council of Medical Research (ICMR) validated it (1). Amidst the chaos and responsibilities of saving the patients’ life this came as a blessing which could give a reliable Point Of Care (POC) test. The test is not only sensitive and specific but also rapid and cheap so it can be used in rural set ups too (2). Gupta N et al., very aptly termed it as “a laboratory in a suitcase” as it is very portable, light weight, battery powered and can be used in areas of low power supply and connectivity (3). Besides, it does not require well equipped laboratory and much manpower compared to other molecular methods. The TrueNat machine, which works on the principle of Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR) has been designed, developed and manufactured by Molbio Diagnostics Private Limited, Goa, India is also equipped for detection of tuberculosis, multidrug resistant tuberculosis, malaria, dengue, Human Immunodeficiency Virus (HIV), Rabies, influenza, etc., (4). Due to its multifaceted properties and uses this chip based rapid test has a promising future in the field of molecular diagnostics.

Thus, this Molbio Quattro (capacity to run 4 samples per run) model was used in the SNM Hospital at Firozabad which is a tertiary care hospital catering to patients from Firozabad district and neighbouring areas. During the pandemic period when it was difficult to establish laboratories for molecular testing of the virus and there were a lot of samples to be tested, the TrueNat testing method came as a blessing and this is the first study from Western Uttar Pradesh, India, to report on its working efficacy. Hence, present study was done to evaluate the importance of TrueNat testing among symptomatic and asymptomatic cases in different age groups and gender and also to see any association of symptoms with E gene and RdRp gene positivity.

Material and Methods

In this retrospective study, which was conducted at Autonomous State Medical College and SNM Hospital, Firozabad, Uttar Pradesh, India from June 2020 - May 2021 (thereafter, the data analysis was done from June-December 2021) wherein, 4,659 samples were tested by the TrueNat machine besides recording the age and gender details. The signs and symptoms were also recorded and the cases were divided into three groups, viz., group A patients who presented with symptoms ≤7 days, group B patients who presented with signs and symptoms >7 days (4) and group C comprising of asymptomatic patients. The data of symptoms and clinical history/travel history was collected from hospital records or patient summary reports.

Inclusion criteria: All the patients of SARI, ILI, symptomatic Outpatient Department (OPD) patients, preoperative, in labour or for caesarean section and emergency or trauma patients (irrespective of being symptomatic or asymptomatic) who required prompt treatment and surgical intervention, contacts and travellers.

Exclusion criteria: Samples from other bacterial respiratory illness like tuberculosis, pneumonia etc., were excluded.

Sample collection and processing: A single oropharyngeal swab was collected by trained staff following the laboratory safety guidelines (5) in VLM which was provided by Molbio company. The machine consists of a nucleic acid extraction device and an automated real time polymerase chain analyser along with accessories like TrueNat SARS CoV-2 micro PCR chips, microtube with freeze dried reagents, RNA cartridges, DNase and RNase free pipette tips, holding stand etc. The kit was stored between 2-30ºC. The RNA was extracted from the patients’ sample using Truepep AUTO V2 Universal Cartridge Based Sample Prep device which is a fully automated device using a fluidic cartridge which extracts the RNA.

Principle of the test- It works on the principle of Real Time RT-PCR. The target sequence for the kit being E gene, RdRp gene and human RNase P which serves as Internal Positive Control (IPC). If sample was positive for Beta CoV, 6 μL of the same extracted RNA was put in the reaction well and the test was inserted in the real time quantitative micro PCR analyser where the RNA is first converted into complementary DNA (cDNA) and further thermal cycling takes place. Positive amplification causes the dual labelled fluorescent probe in the chip based Real Time PCR test to release fluorophores in an exponential manner which is captured by the built in optoelectronic sensor and displayed on the screen.

Interpretation of result: The results are obtained as amplification curves. Both the target and IPC curves take an exponential path and the fluorescence crosses the threshold value in case of positive samples. The curve remains horizontal throughout the test and the IPC curve takes an exponential path in case of negative samples. In case IPC remains horizontal in a negative sample the test is invalid (displayed on the screen).

At the end of the test, results are displayed as not detected for negative and detected for positive results. In case of positive results, the viral load is displayed as High (Ct<20), Medium (20≤Ct<25), Low (25≤Ct<30) and Very low (Ct ≥30) as per the manufacturers’ instructions. The Ct values for positive samples were recorded and association with the duration of symptoms in the patients.

Statistical Analysis

All the variables were presented in the form of frequencies and percentages besides being depicted in suitable diagrammatical representation for the bird-eye-view. Thereafter, the Chi-square test was done to test the statistical significance of association of symptomatic patients with the outcome of the TrueNat test. The association of categories of Ct values and symptomatic patient was tabulated and its significance checked using Chi-square test. All the analyses were performed using R-3.6.2 and MS Excel 2007.

Results

Of the total samples (4659) tested, the maximum (1169) testing for COVID-19 was done in the females of the age group 20-29 years followed by 30-39 years. In other age groups TrueNat testing was predominantly done on males (Table/Fig 1). The maximum number of positive cases (72) were found in people of age groups 20-39 years and least positivity occurred in age groups below 9 years (two cases) and above 80 years (two cases) (Table/Fig 2).

The positivity percentage detected by TrueNat testing was 3.3%. There were nine samples which gave invalid results and had to be repeated. Of the total number of people tested for travel purpose (590 cases), the maximum positive cases were seen in the age group 20-29. The contacts of positive cases showed maximum positivity 2.2% (Table/Fig 3). The maximum TrueNat positivity was seen in patients who presented with symptoms <7 days and on association with the Ct values of E gene and RdRp gene it was found that 28.1% and 27.2% of these patients had high Ct values group (Table/Fig 4). Of the total 153 positive samples, 147 samples showed both E gene and RdRp gene positivity and six samples showed presence of only E gene. There were 21 cases in which the E gene as well as the confirmatory RdRp gene was detected but the patients were asymptomatic.

Discussion

The emergence of the COVID-19 pandemic gave rise to increased testing by different methods. Also, the guidelines laid down by the WHO necessitated the screening of people so that they could be sent to isolation timely to stop further spread of the infection (6). In the present study, the maximum TrueNat testing was done in females of the age group of 20-39 years. This being the reproductive age group, these women approached the hospital either for deliveries or other surgeries. It was followed by testing in males of age groups above 40 years. This was because the locomotion of males due to work and need of testing before national and international travelling. The positivity rate was 3.3% in the present study and maximum incidence of infection was found in the age groups 20-29 years followed by 30-39 years (p<0.05). Bharti S et al., in their study also observed maximum COVID-19 infection in the age groups 26-30 years (7). Though in countries like Italy there was a sharp contrast where 69% of infected people were in the age groups 51-70 years (8). There was no significant impact on positivity on the basis of gender in the present study. However, Bharti S et al., observed that females were affected less by COVID-19 compared to males (7) and their results corroborated with worldwide estimates where the authors found that the vulnerability of the males to infection was 1.14 times compared to females (9). This could be due to more locomotion and social interaction of males. In the present study, the people who got tested for traveling purpose were found positive maximum in the age group of 20-29 years which is an age of more locomotion due to professional reasons.

In the present study, patients who presented with symptoms for <7 days had high Ct values (<20) for E gene and RdRp gene. In patients presenting with symptoms >7 days there were more samples showing Ct values in the medium, low and very low range. The association of Ct value and the duration of symptoms showed that the longer the duration of infection the lower the Ct value (p<0.05) and vice versa. The Ct value is the number of amplification cycles needed to produce a fluorescent signal (10). Thus, the low Ct value (numerical) indicated high viral RNA load (11). However, the Ct values do not have a direct association with the disease severity and could be inversely proportional to the viral load and transmissibility (12),(13). In a retrospective study conducted by Shah S et al., they did not find any association between severity of disease and the Ct values (14). Aranha C et al., have shown that the viral RNA detection by molecular techniques does not determine the infectivity of the virus or presence of replicative virus (15). Many studies have shown that the high Ct values (numerical) correspond to non infectious viral RNA determined by viral culture (16),(17),(18). Laferl H et al., concluded in their study that the samples with Ct values>30 corresponded to non viable particles that could be still detected by molecular methods (19). In that study, people presenting with the symptoms for more than a week also showed high Ct values (E gene- 14.4% patients, RdRp- 13.6% patients). This could be due to their decreased immunity or comorbid conditions which delayed recovery. However, there have been studies which haven’t studied the disease severity with the Ct values (20),(21). Similarly, there have been studies that have not found difference between viral loads as determined by Ct values between symptomatic and asymptomatic patients (22). In the present study, six samples which showed the presence of only E gene and the absence of the confirmatory RdRp gene, patients could be suffering from viral infection due to some other member of the Coronaviridae family. Studies have shown a significant homology of the E gene to other Coronaviruses (23). There were 21 asymptomatic cases which showed the presence of E gene as well as RdRp gene. Singanayagam A et al., also concluded that asymptomatic people represented as a source of potential transmissible virus (24). These people would have acquired the infection sub clinically but owing to good immunity evaded the symptoms. Since, the virus detected in the samples of the low Ct value group does not predict infectivity of the person, these could be from the non replicating virus indicating infection in the near past or the patients could be in convalescence stage. Asai N et al., concluded that the Ct values of molecular tests decreased with patients’ recovery and in some asymptomatic patients these were positive even for longer than two weeks (25). The molecular detection does not differentiate between infectious and non infectious virus (24).

The TrueNat method being a molecular technique could detect the viral RNA even in traces. The advantage of TrueNat over the conventional rRT-PCR method was that the time period of RNA extraction was reduced to less than 60 minutes (26). This is because the machine uses a disposable fluidic cartridge to extract RNA from the VLM in 15 minutes. This advantage of TrueNat made it a very patient friendly technique, time being of utmost importance in the detection of COVID-19 virus so as to manage patients effectively by facilitating isolation. Gibani MM et al., while assessing the COVID Nudge, a POC test also found it reliable, sensitive 100%, specific 94% with a turnaround time of 90 minutes per test (27). Since the step of manual RNA extraction was omitted, it reduced the risk of contamination and thus reduction in false positive results. The other advantage of the VLM being, it lyses the microorganisms rendering them non infectious thus, offering protection and making it user friendly. In a study conducted by Erster O et al., they found that using lysis medium over Viral Transport Medium (VTM) increased the sensitivity, safety and rapidity of COVID-19 testing and also allowed sample preservation for longer period without any special cooling equipment (28). Ghoshal U et al., estimated the cost per test to be only 15 USD which is cost effective (29). Besides the machine does not require much space (can be kept on a table top) or air conditioned laboratory which not only makes it more cost effective but also adjustable anywhere. It is not very labour intensive. The battery is rechargeable. Being a Make in India technology it has been very prestigious to have an indigenous diagnostic product in the country at the time of pandemic (30).

Limitation(s)

The limitation was in getting proper clinical history in some cases due to the fear of interaction with COVID-19 symptomatic patients and while performing the tests few samples had to be repeated due to invalid results.

Conclusion

Thus, the TrueNat method of detecting the COVID-19 virus is fast, easy to perform and can also be used in rural set ups. Since, it is a molecular method, it can be used for the confirmed diagnosis of the COVID-19 virus and is very helpful in set ups that do not have the facility to perform rRT-PCR. It can be used effectively as an epidemiological tool. The Ct values should be considered by the clinicians and correlated with the symptoms. In this retrospective study, the E gene could be detected in few samples but not the confirmatory gene, indicating the presence of infection due to any other member of the Coronaviridae family and an added advantage to the technique.

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DOI and Others

DOI: 10.7860/JCDR/2022/55742.16292

Date of Submission: Feb 16, 2022
Date of Peer Review: Mar 03, 2022
Date of Acceptance: Mar 14, 2022
Date of Publishing: May 01, 2022

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? NA
• Was informed consent obtained from the subjects involved in the study? NA
• For any images presented appropriate consent has been obtained from the subjects. NA

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• Plagiarism X-checker: Feb 23, 2022
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• iThenticate Software: Mar 12, 2022 (9%)

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