Sonali Bhattar

Rapid Tests versus ELISA for Screening of HIV Infection: Our Experience from a Voluntary Counselling and Testing Facility of a Tertiary Care Centre in North India

Early and accurate diagnosis of human immunodeficiency virus (HIV) infection is essential for timely identification of patients needing antiretroviral therapy and for instituting HIV prevention strategies. The primary methodology for HIV testing has shifted from enzyme linked immunosorbent assay (ELISA) to rapid diagnostic tests (RDTs) in recent years, especially in resource limited settings. However, the diagnostic performance of RDTs is a matter of concern. In the present study the performance of an RDT being used as the initial test in serial testing based algorithm for HIV diagnosis was compared with ELISA. Seven hundred and eighty-seven sera, tested at the voluntary counselling and testing facility employing a serial testing algorithm (based on SD Bioline HIV-1/2 3.0 as the first test), were subsequently tested with Microlisa-HIV for anti-HIV antibodies. The first test missed 9 HIV reactive samples and also registered 5 false positives. The sensitivity, specificity, and negative and positive predictive values of the first test were 77.5%, 99.3%, and 98.8% and 86.1%, respectively, taking ELISA as the standard test. Our study highlights that RDTs fare poorly compared to ELISA as screening assays and that reactive results by RDTs need to be confirmed by western blot for a positive serodiagnosis of HIV infection.

Evaluation of SD BIOLINE Syphilis 3.0 for rapid diagnosis of syphilis: Report from a regional sexually transmitted infection reference laboratory in North India

Background: Serology is considered the mainstay of syphilis diagnosis. The limitations of the traditional serological methods and the advent and availability of novel immunochromatographic assays have led to the widespread application of rapid point-of-care procedures as screening tools for syphilis. However, these tests have not been extensively evaluated. This study was designed to evaluate the performance of a rapid syphilis diagnostic test known as SD BIOLINE Syphilis 3.0 (SD Biostandard Diagnostics Private Limited, Gurgaon, Haryana, India). Materials and Methods: A panel comprising of 50 venereal disease research laboratory reactive and 50 nonreactive sera was tested using SD BIOLINE Syphilis 3.0. The performance of the test was evaluated using IMMUTREP Treponema pallidum hemagglutination assay (TPHA) (OMEGA Diagnostics Limited, Scotland, United Kingdom) as the reference standard and sensitivity, specificity, and negative and positive predictive values were calculated. Results: The sensitivity, specificity, and positive and negative predictive values of SD BIOLINE Syphilis 3.0 were 92.86% (confidence interval of 95%: 80.52–98.50%), 98.28% (90.76-99.96%), 97.50% (86.84–99.94%), and 95.00% (86.08–98.96%), respectively, compared to TPHA as the gold standard. Conclusion: Keeping in view the high sensitivity and specificity of SD BIOLINE Syphilis 3.0, we conclude that the test can be used as a tool for rapid on-site diagnosis of syphilis and as an alternative to TPHA for detection of antibodies to Treponema pallidum.

Sexually transmitted infections and HIV in self reporting men who have sex with men: A two-year study from India

Homosexuality is not legally and socially accepted in India. Thus, this area of research has largely been ignored by Indian authors, resulting in dearth of knowledge, particularly with respect to sexually transmitted infections (STIs) in this high-risk group. Over a period of two years (2013-2014), 738 males sought care at skin and venereal diseases clinics, 52 (7.05%, 95% CI=5.4-9.14%) of who identified themselves as MSM and were enrolled in the study. Diagnosis was made on the basis of clinical presentation and laboratory testing, wherever indicated. Thirty six percent of MSM had only homosexual preferences, while 64% were bisexual. The most common sexually transmitted infection was genital warts (23.08%, 95% CI=13.58-36.28%). Fourteen patients (26.92%, 95% CI=16.67-40.35%) were VDRL and TPHA positive (two, five and four with primary syphilis, secondary syphilis and latent syphilis, respectively). These were followed by genital herpes (11.54%, 95% CI=5.03-23.34%), genital molluscum contagiosum (9.62% 95% CI=3.75-21.04%), and gonorrhea (5.77%, 95% CI=1.38-16.25%). Of those tested, 23.08% (95% CI=13.58-36.28%) of patients were reactive for HIV serology. Thus, MSM is a high-risk group with high prevalence of HIV and other STIs in this group, mandating greater focus, education and counseling.

HIV/AIDS Awareness among VCT Clients: A Cross-Sectional Study from Delhi, India

The contribution of India to the global burden of HIV/AIDS is significant. A major barrier that the country has faced in its battle against this disease is the inadequate and inaccurate information about it among the population. The present analysis explores the knowledge about HIV/AIDS among clients attending a voluntary counselling and testing (VCT) facility in India. Two hundred clients attending the VCT facility were assessed in this regard using a structured predesigned questionnaire. Sixty-three (31.5%) of the respondents had never heard of HIV/AIDS. In comparison to males, a significantly higher number of females had not heard about the disease (P < 0.01). Lower levels of education of participants were found to be significantly associated with the response of not having heard of HIV/AIDS (P < 0.01) as was an occupation status of being an unemployed man/housewife (P = 0.002). For the 137 (68.5%) respondents who had heard about HIV/AIDS, television was the source of information in 130 (94.9%) followed by posters in 93 (67.9%) and newspapers in 88 (64.2%). While the knowledge about HIV transmission and prevention was good, the extent of misconceptions was high (61.8%). Our study highlights the strong need to raise the levels of HIV awareness among Indian population.

Chlamydia trachomatis Infection in HIV-Infected Women: Need for Screening by a Sensitive and Specific Test

Reproductive tract infection (RTIs)/sexually transmitted infections (STIs) are recognized as a major public health problem, particularly due to their relationship with HIV infection. Early detection and treatment of Chlamydia trachomatis infection (CTI) among HIV-infected and HIV-uninfected women may impact heterosexual HIV transmission. A total of 120 participants were enrolled: 30 HIV seropositive women with symptoms of RTIs, 30 HIV seropositive women without symptoms of RTIs, 30 HIV seronegative women with symptoms of RTIs, and 30 HIV seronegative women without symptoms of RTIs. One endocervical swab was collected from all participants and CTI was detected by real-time PCR (COBAS TaqMan CT Test, v2.0). CTI was detected in 4 (6.67%) HIV-infected women and in 1 (1.67%) HIV-uninfected woman (OR 4.214; 95% CI 0.457–38.865). Vaginal discharge was present in almost half of HIV-infected and HIV-uninfected women; lower abdominal pain was present in 11 (18.3%) of HIV-infected and in 9 (15%) of HIV-uninfected women. This study showed that CTI is more prevalent among HIV-infected females as compared to HIV-uninfected females. As the use of real-time PCR is not feasible in most hospitals, efforts should be made to develop a simple, sensitive, and specific test to identify women with CTI for prevention of sequelae and HIV transmission.

Correlation of CD4 T Cell Count and Plasma Viral Load with Reproductive Tract Infections/Sexually Transmitted Infections in HIV Infected Females

BACKGROUND Sexually transmitted infections (STIs) plays a major role in the spread of Human immunodeficiency virus (HIV) due to common route of transmission. These infections display an epidemiological synergy with HIV. AIM The aim of this study was to analyse the correlation of CD4 T lymphocyte cell count, HIV-1 plasma viral load with Reproductive tract infections/Sexually transmitted infections (RTIs/STIs) in HIV infected females. MATERIALS AND METHODS The study included 60 HIV infected females. An informed consent was taken from all the study subjects. Relevant specimens (genital specimen and blood) were collected for laboratory diagnosis of various RTIs/STIs, CD4 cell count and plasma viral load estimation. RESULTS Mean CD4 count of females with bacterial vaginosis, vaginal candidiasis, trichomoniasis, syphilis and herpes simplex infection were lower as compared to other HIV infected cases and mean plasma viral load of bacterial vaginosis, vaginal candidiasis, trichomoniasis and syphilis were higher as compared to other HIV infected cases but this difference was not statistically significant. CONCLUSION This study highlights the importance of routine screening for STIs/RTIs of all the HIV infected females for RTIs/STIs irrespective of CD4 cell count and plasma viral load.

Correlation between venereal diseases research laboratory titers and CD4 T-lymphocyte count determined by flow cytometry in HIV-infected adults: A 5-year study

Introduction: Venereal Diseases Research Laboratory (VDRL) is one of the key tests for the diagnosis of syphilis; however in HIV-positive individuals, it has been reported to give inappropriate results at times. Thus, this study was conducted to determine if the VDRL test titers vary with the severity of immunosuppression as determined by CD4 cell count. Materials and Methods: A total of 2630 samples from HIV-positive adults were tested by qualitative and quantitative VDRL test and if reactive, by Treponema pallidum hemagglutination (TPHA) test. CD4 cell counts were determined at the same time by flow cytometry (BD FACSCount™ system). Correlation between CD4 T-lymphocyte cell count and VDRL titers was sought for. Results: Nearly 6.7% (176/2631) of individuals were VDRL reactive, males more than females (7.6% vs. 5.1%, P = 0.014). Four of the VDRL-reactive patients were found negative by TPHA test and were excluded from further study. VDRL titers ranged from weakly reactive to being reactive at 1:128 (median = 1:2). The CD4 cell count ranged from 23 cells/μl to 883 cells/μl (median = 276 cells/μl, mean = 323.9 ± 200.9). Pearsons coefficient of correlation (R) between CD4 cell count and VDRL titers was calculated to be 0.0559; coefficient of determination (R2) was 0.0031. Conclusions: Although the correlation coefficient shows a positive correlation, the association was very weak. Therefore, CD4 cell count cannot be expected to influence VDRL titers in HIV-positive adults significantly.

Evaluation of Baseline CD4+ T Cell Counts and ART Requirement in Newly Diagnosed HIV Seropositive Individuals in a Tertiary Care Hospital of Northern India.

INTRODUCTION Antiretroviral Therapy (ART) has changed the outlook of Human Immune-deficiency Virus (HIV)/Acquired Immuno Deficiency Syndrome (AIDS) patients worldwide. AIM To analyse the trends in baseline CD4+ T cell counts and ART requirements in newly diagnosed HIV seropositive individuals in a Tertiary care hospital of Northern India. MATERIALS AND METHODS Out of 1263 HIV seropositive clients identified from January 2012 to June 2014, the baseline CD4+ T cell counts of only those 470 clients were analysed, who registered at the linked ART centre. RESULTS The mean baseline CD4+ count of the study group was 249.77±216.0cells/mm3 and that of male and female were 300.31±240.47cells/mm3 and 232.38±204.25cells/mm3 respectively. A total of 259 of 334 (77.54%) HIV reactive males, 83 of 130 (63.85%) HIV reactive females and overall 348 of 470 (74.04%) required antiretroviral treatment on enrolment. CONCLUSION In the present study, about three-fourth of newly diagnosed HIV positive Indian patients required initiation of ART at registration. The relatively low baseline CD4+ T cell counts in this population highlights the need for timely baseline CD4+ counts testing of HIV positive patients and the urgency of initiating treatment in HIV reactive individuals in Indian health care settings.

Utility of Laboratory Diagnosis for Confirmation of the Syndromic Case Management in Married Indian Women with Vaginal Discharge

OBJECTIVE WHO and NACO recommend treatment of STIs/RTIs on the basis syndromic case management (SCM), even without laboratory confirmation, which may lead to over-treatment. Thus, this study was conducted to evaluate the utility of laboratory diagnosis for confirmation of patients with vaginal discharge diagnosed on the basis of SCM. METHODOLOGY 234 married women in reproductive age group, diagnosed as having vaginal discharge syndrome were included. Normal saline wet-mount slide preparations were made for detection of motile trichomonads. Gram stained smear were prepared and scored as per classification developed by Nugent. The presence of pseudohyphae and/or budding yeast cells was considered diagnostic of candidal infection. VDRL, TPHA and HIV testing were also done as per protocol. RESULTS The median age of the study population was 34 years. Most common cause was bacterial vaginosis (positive= 21.4%, 95% CI= 16.6-27.1%; intermediate score= 17.5%, 95% CI= 13.2-22.9%), followed by candidiasis (13.7%, 95% CI= 98-18.7%) and trichomoniasis (0.4%, 95% CI= 0-2.6%). No etiological diagnosis for vaginal discharge could be established in approximately half of the women. Only two women were HIV positive; one was reactive by VDRL and TPHA tests. CONCLUSION Our study highlights the possible lacunae in SCM. Large number of patients may be over-treated if only syndromic management is followed, with financial, medical and social implications. Thus we recommend, the treatment maybe initiated on the basis of SCM, but it is essential that laboratory diagnosis is sought for and the treatment modified accordingly.

Seroprevalence of syphilis by VDRL test and biological false positive reactions in different patient populations: Is it alarming? Our experience from a tertiary care center in India

Introduction: Many centers for sexually transmitted infections in India perform only a single screening assay for diagnosis of syphilis which may yield biological false positive (BFP) reactions. Aims and Objective: The aim of this study was to determine the true picture of seroprevalence of syphilis and BFP reactions in different patient groups. Materials and Methods: A total of 57,308 serial serum samples obtained over a period of 5 years from different patient groups were screened by venereal disease research laboratory (VDRL) test both qualitatively and quantitatively. VDRL reactive sera were confirmed by Treponema pallidum hemagglutination (TPHA) test. Results: The overall seroprevalence of syphilis by VDRL test was 1.27%, and BFP rate in test population was 0.14%. The rate of BFP reactions among total tested male (0.44%) and female (0.1%) patients differs significantly. Out of 733 VDRL reactive samples, 81 were BFP, i.e., BFP reaction is occurring at a frequency of 11% of the total VDRL reactive samples (ratio of 8:1 for true positives/BFP). Similarly, among antenatal cases, almost 24% of the total VDRL reactive samples were BFP, or for every 116 true positives, there were 37 (almost one-third) BFP. Conclusion: Although the overall seroprevalence of syphilis is low; the frequency of occurrence of BFP reactions is quite alarming. Hence, treponemal test must be used for confirmation of VDRL reactive sera.