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

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Dr Mohan Z Mani

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Believers Church Medical College,
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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.
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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 : February | Volume : 16 | Issue : 2 | Page : GC01 - GC04 Full Version

Sperm DNA Fragmentation Index in Infertility Patients vs Normal Individuals: A Cross-sectional Study


Published: February 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/51563.15979
Manisha B Sinha, Nighat Hussain, Sarita Agrawal

1. Additional Professor, Department of Anatomy, AIIMS, Raipur, Chattisgarh, India. 2. Professor, Department of Pathology, AIIMS, Raipur, Chattisgarh, India. 3. Professor, Department of Obstetrics and Gynaecology, AIIMS, Raipur, Chattisgarh, India

Correspondence Address :
Dr. Manisha B Sinha,
Additional Professor, Department of Anatomy, AIIMS Medical College, Gate No-5,
Raipur, Chattishgarh, India.
E-mail: manishab80@gmail.com

Abstract

Introduction: Infertility is a major problem among married couples. The male factor contributes to around 30-40% of all cases of infertility. With advancing age, reducing sperm count may magnify the problem. The awareness of qualitative change along with quantity is important for people seeking advice in infertility.

Aim: To find the prevalence of sperm Deoxyribonucleic Acid (DNA) fragmentation with Sperm Chromatin Dispersion (SCD) test.

Materials and Methods: The present study was prospective cross-sectional study which was carried out on semen sample given for routine check-up in department during the period from August 2017 to August 2018. A total of 180 couples, including 88 infertile couples and 92 couples with known fertility were enrolled in the study. Sperm Count and sperm DNA Fragmentation Index (DFI) were calculated and determined by SCD test by kit method. Comparison between groups was done according to distribution of data using Mann-Whitney rank-sum (two tailed) statistical test.

Results: Among 180 subjects, (88 cases and 92 control) no significant difference in the age was found. The sperm count was not significantly different in cases (group of men of infertile couple) and controls (group of men of proven fertility). The highest value for DFI was 27% in controls and 97% in cases. Statistically significant difference was found in DFI (p=0.008) in both groups. DFI and sperm count showed negative correlation both in cases and controls with a correlation coefficient 0.213 (p-value <0.001) and 0.754 (p-value <0.005) respectively. A significant difference was observed in the median value of sperm DFI of men of infertile couple as compared to men of fertile couple.

Conclusion: On comparing, significant difference was observed in the median value of sperm DFI in infertile couple. Sperm DFI at cut-off value 27% has higher diagnostic significance. SCD method is simple, easiest and standard tool to assess DFI.

Keywords

Deoxyribonucleic acid, Normozoospermia, Oligozoospermic, Sperm chromatin dispersion test, Sperm count

Infertility is an important and common problem in our society. This is also quite common in affluent and learned group. This problem affects men and women equally. Out of total males, every six men in our society are infertile, which is quiet a depressing figure (1),(2),(3). The problem becomes more, as many men may not accept infertility.

To diagnose infertility due to male factor, the sperm count is an important but not a sufficient test. Once sperm count is reported normal, a man feels happy that he has no problem. However, it’s not so because the quality of sperm is also very important to give positive outcome in the form of pregnancy. To test sperm parameters, usually a doctor asks for sperm count, morphology and motility. However, to test the quality of sperm, it is also worthwhile to check for sperm DNA fragmentation (SDF). The SDF reflects the integrity of genetic material of the gamete (4). The intact sperm DNA in turn is responsible for zygote integrity and pregnancy outcome. SDF can be a result of many factors: smoking, alcoholism, high local temperature, varicocele etc. Some of them are modifiable by adding antioxidants and astaxanthins (5). Hence, identification of SDF, prior counseling and appropriate interventions may help in improving the fertility outcome (6).

In last three decades, many techniques have been developed to evaluate sperm chromatin quality in reproductive molecular biology (7),(8). The sperm chromatin quality is assessed in the form of sperm DFI. In a country like India, people try to avoid expensive test. DFI can be assessed by Sperm Chromatin Structure Assay (SCSA), TUNEL assay (TdT-mediated –dUTP nick end labelling), COMET assay (single gel electrophoresis) and Acridine Orange-staining Technique (AOT). The newer and economic technique is SCD test. This assay is based on the principle that sperm with fragmented DNA fail to produce characteristic halo of dispersion that is observed in sperm with non fragmented DNA following acid denaturation (9).

Many studies have concluded that sperm DNA fragmentation as one of the factor responsible of male factor infertility (10),(11). However, it is not yet incorporated or recommended as a part of routine investigation of infertility. The aim of present study was to find the prevalence of sperm DNA fragmentation in semen sample of male partner of infertile couple and its correlation with conventional semen parameters with special reference to sperm count in the central India population.

Material and Methods

The present study was a prospective cross-sectional study in which 88 infertile couples visiting infertility clinic in Department of Obstetrics and Gynaecology, AIIMS, Raipur, Chattishgarh, India, and 92 couples with known fertility were included. Study was conducted from August 2017 to October 2018. Institutional Ethical Clearance (IEC) (AIIMSRPR/IEC/2016/046) was obtained.

Inclusion criteria: Normozoospermic infertile couple, Oligozoosp-ermic infertile couple in age group 22-45 years were included as a case. For the comparable control, age was same and either normozoospermia or oligozoospermic having child.

Exclusion criteria: Men with history of testicular maldescent, testicular malignancy or testicular injury including torsion were excluded.

Study Procedure

Eighty-eight men from infertile couples and 92 men from couples with known fertility were enrolled in the study. Men from infertile couples were taken as cases and men with known fertility were taken as control group. Patient information sheets were given to all and explained about the outcome. After obtaining written consent for participation, patients were asked to give semen sample usually after three days of abstinence. Each sample was allowed to liquefy for 20-30 minutes at 37°C. After evaluating volume, samples were aliquoted in two parts. One part of the sample was used to evaluate basic sperm parameters including sperm count as per to World Health Organisation (WHO) guideline (12) in Department of Pathology and Laboratory Medicine. Based on the total sperm count, the cases were divided in two subgroups; normozoospermic (>15 million/mL) and oligozoopermic group (<15 million/mL).

The second part was used to assess sperm DNA fragmentation in Department of Anatomy. The sperm DNA fragmentation was carried out by SCD test: Sperm DNA fragmentation test was done by kit sperm 360 DNA fragmentation (Sperm Processor Pvt. Ltd., Aurangabad, India). This kit is based on protocol illustrated by Fernandez JL et al., and further modified by Chauhan KR et al., using 0.4 M Dithiothreitol (DTT) (9),(13). An aliquot of semen sample is mixed with 1% low melting agarose at 37°C. Aliquots of 60 μL of the mixture were pipetted and spread on slide and covered with coverslip. Slides were left to solidify at 4°C. After that the slide was treated with reagent I (lysis solution) for seven minutes. Then the slide was treated with reagent II (neutralising and lysis solution 2) then washed with distilled water. The slide was then dehydrated sequentially in 70%, 90% and 100% ethanol, keeping for two minutes in each. Slide was stained with stain provided in kit for microscopy. Thereafter, the slide was examined under bright field microscope. On each slide, 500 sperms were evaluated for halo size and dispersion pattern as described by Fernandez JL et al., (9) for: 1) nuclei with large DNA dispersion halos; 2) nuclei with medium sized halos; 3) nuclei with small sized halos; and 4) nuclei with no halo. The nuclei with large to medium size halo were considered as non fragmented DNA (dispersed nuclei). The nuclei with small size halo and no halo at all were considered as fragmented DNA (non dispersed nuclei) (Table/Fig 1). DFI were calculated as percentage of number of spermatozoa with fragmented DNA out of number of spermatozoa counted. The subjects were further divided according to DFI with cut-off values 25%, 15%-25% and <15% (14).

Statistical Analysis

Statistical analysis was done using Statistical Package for the Social Sciences (SPSS) (version 21.0) software. Comparison between groups was done for values of Body Mass Index (BMI), sperm count, DFI by using Mann-Whitney rank-sum (two tailed) statistical test. Results were expressed in mean±Standard Deviation (SD). Percentage of non fragmented DNA/dispersed nuclei, fragmented DNA/non dispersed nuclei and dead nuclei were calculated in cases and control. Pearson’s correlation test was used to correlate various variables in the study. The p-value of <0.05 was considered as statistically significant.

Results

All variables tested by Mann-Whitney rank-sum test were found to be variably distributed. Results of parameters measured in the study are shown in (Table/Fig 2). Demographic profiles were not significantly different. The cases showed decreased sperm count as compared to controls. However, this was not statistically significant. Percentage of non fragmented sperm count were significantly lower in cases than in control (p= 0.011). Percentage or fragmented sperm and dead sperm were significantly higher than control (p=0.026, p=0.05, respectively).

The highest value of DFI in cases was 93% as compared to 27% in controls. This was statistically significant (p=0.008). On comparing DFI value was significantly high in men from control group as compared to men from infertile couple.

DFI and sperm count showed negative correlation both in cases and control with a correlation coefficient being 0.213 (p-value <0.01) and 0.754 (p-value <0.005), respectively. Median value of sperm DFI in cases was significantly higher, 23% (3-93%), compared to 10.5% (3-27%) {(Table/Fig 3) box plot}.

The discriminating power of DFI measured by SCD test, to identify threshold value between cases and control, was calculated by ROC curve analyses. During ROC analyses (Table/Fig 4), varying percentage of DFI values was used to calculate optimum sensitivity and specificity value for SCD test. The best area under cover ROC curve was 0.770 for 27% of DFI (Table/Fig 4). With the cut-off point 27%, sperm DFI was able to distinguish cases and control. With this threshold had 98.91% (94.01.52% to 99.97%) specificity and sensitivity of 43.18% (95% CI 32.66 to 54.18). The Positive Predictive Value (PPV) and Negative Predictive Value (NPV) of the 27% sperm DFI were 97.44% (95% CI 84.21 to 99.63) and 64.54 (95% CI 60.24 to 68.62%), respectively. The sperm DFI value >27% was found in 31% (20/65) of normozoospermic cases and 78% (18/23) of oligozoospermic cases (Table/Fig 5).

Discussion

In spite of the periodic refinements in semen analysis techniques and cut-off values in the form of WHO guidelines for semen assessment, upto 30% of men with normal semen parameters remain infertile (15). The studies advocate testing the sperm DNA fragmentation test before going for Assisted Reproductive Technology (ART). In order to avoid an increase in budget, whom to recommend should be assessed. Studies suggest that sperm DNA fragmentation test appears to have strong correlation with, In Vitro Fertilisation (IVF) and Intracytoplasmic Sperm Injection (ICSI) outcome (16),(17).

In present study, there was no significant difference in the age and BMI of two groups (Table/Fig 1). Studies have reported that ageing decreases the function of organs hence the sperm count. With increasing infertility problem, one should go for DNA fragmentation test without any delay. Frattareli JL et al., found age related decrease in ability of spermatozoa to fertilise ova (18). Other studies reported that age has no effect on fertilisation rate (19) and still others have shown negative correlation with paternal age for ICSI (20).

In current study, DFI value was significantly higher in men from infertile couple as compared to men from control group which suggests high DNA fragmentation is associated with infertility. Threshold DFI value (27%) showing infertility in the current study was comparable to earlier study done by Larson KL et al., (21). They had also reported absence of clinical pregnancy at or above DFI threshold value of 27%. Also, in their study using these samples in ART i.e., IVF and ICSI cycles did not give good results. Fernandez JL et al., found statistically significant difference in sperm DFI values in healthy sperm donor and infertility patients (16.7±9.9 Vs 35.4±18.3, p<0.05), tested with SCD test (9). Wiweko B and Utami P in their study again found significant difference in DFI in healthy fertile men and infertile men (19.9% vs 29.9%, p<0.001) (22). Sperm nuclear protein, the protamine and Transitional Protein (TP), are responsible for integrity of compacting of sperm DNA. Sperm nuclear protein plays vital role in integrity of compact sperm DNA during spermatogenesis (23),(24),(25),(26),(27). Sperm DNA damage is because of protamine deficiency. In the current study, median value of DFI was significantly higher in cases as compared to control. Wiweko B and Utami P also found similar results (22).

In the current study, 32.3% of normozoospermic cases and 83% of oligozoospermic cases had sperm DFI value >25%. In the current study, 31% of normozoospermic cases and 78% of oligozoospermic cases have sperm DFI value >27%. It means the probability of oligozoospermic cases to have high sperm DFI value was significantly higher as compared to normozoospermic cases. However, Fernandez JL et al., found no statistically significant differences in sperm DFI values from infertility patients with normal or abnormal semen parameters (32.1± 20.4 vs 38.7±16.3, p>0.05) (9).

Correlation between DFI and semen parameters was found to be only weak-to moderate (r=0.213). This was statistically significant (χ2 value=1.67, p-value=0.046). Normozoospermic cases and control also had significant difference in preponderance of DFI (χ2 value=17.67, p-value=0.001). Other studies also showed similar finding (28),(29),(30).

In current study, the method adopted to study the sperm DNA fragmentation was SCD. In this, on agarose fixed sperm, acid denaturation solution is used to segregate single stranded DNA (31) and is responsible for suppression of production of DNA halo. Treatment with lysis solution is responsible for disintegration of single stranded DNA and non formation of DNA halo in fragmented sperm. The result of ROC curve analysis indicated that sperm DNA fragmentation, as measured by SCD test, was a good predictive parameter to distinguish between fertile and infertile population on the basis of largest AUC {0.77; [Table/Fig- 2]} (p<0.002; 95% CI 0.656-0.885). Wiweko B and Utami P also found AUC value 0.862 (p<0.001; 95% CI 0.783-0.941) (22).

Various studies have produced correlation between sperm DFI value and clinical pregnancy outcome. Meta-analysis and systemic review by Zini A, Osman A et al., Agarwal A et al., have evidenced a significant correlation between sperm DFI value and pregnancy rates with IUI and IVF {Odds ratio (OR)= 9.9, p<0.001} (32),(33),(34). Additionally, Zini A et al., reported correlation between sperm DFI value and miscarriage rate after IVF and ICSI (combined OR=2.48, p<0.0001) (35).

Limitation(s)

Limitations of present study design are: 1) A very small sample size was enrolled in the study; 2) Participants were selected from one Institute and therefore may not be representative of all patients with sperm defragmentation in general central Indian population. Many other environmental factors may contribute to sperm DNA damage in different parts of India.

Conclusion

On comparing, significant difference was observed in the median value of sperm DFI of men of infertile couple. The SCD test is a simpler method for the analysis of sperm DNA fragmentation in semen. This test offers simple and reliable way to screen intact sperm, sperm quality. In the management of infertility patients, the inclusion of SCD test along with the routine semen analysis may help estimate the success of ART. Thus, it is suggested that sperm DFI with a cut-off of 27% by SCD method may be used along with semen analysis before sending an infertile couple for ART in this population.

Acknowledgement

Authors would like to thank Dr. Arvind, CFM, AIIMS, Raipur for statistics calculation.

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DOI and Others

DOI: 10.7860/JCDR/2022/51563.15979

Date of Submission: Jul 25, 2021
Date of Peer Review: Sep 28, 2021
Date of Acceptance: Jan 04, 2022
Date of Publishing: Feb 01, 2022

AUTHOR DECLARATION:
• Financial or Other Competing Interests: This study was financially supported by Institute grant.
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? Yes
• For any images presented appropriate consent has been obtained from the subjects. Yes

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Jul 26, 2021
• Manual Googling: Dec 22, 2021
• iThenticate Software: Jan 03, 2022 (14%)

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