Lyme Borreliosis, a public health concern in India: Findings of Borrelia burgdorferi serosurvey from two states.

Background: Lyme borreliosis is a public health concern in India. The prevalence of the disease is still undetermined with major entomological and epidemiological gaps. The present study was conducted to determine the seropositivity of Borrelia burgdorferi in Sikkim and Arunachal Pradesh, India. Methods: A cross-sectional serosurvey was conducted in Sikkim and Arunachal Pradesh. Data collection tools were developed and standardized for the collection of clinico-socio-demographic data. Sample size for each site was calculated using the formula for the estimation of a single proportion. Qualitative detection of IgG antibodies in serum samples was done using NovaLisa™ Lyme Borrelia IgG ELISA kit. Results: A total of 793 participants were enrolled, 484 (61%) from Arunachal Pradesh and 309 (39%) from Sikkim. Out of 793 participants, 21 (2.7%), 22 (2.8%), 6 (0.8%), 29 (3.7%), 44 (5.5%), and 16 (2.1%) gave history of tick bite, rash, erythema migrans, migratory muscle pain, migratory joint pain, and numbness, respectively, in the past one year. The adjusted seroprevalence (for sensitivity and specificity of kit) for the study is 3.7 (2.4-5.2). No signs or symptoms were found to be associated with IgG ELISA positivity. The state-wise distribution of seropositivity for Arunachal Pradesh and Sikkim was 4.1 (95% CI: 2.5-6.3) and 2.3 (95% CI: 0.9-4.6), respectively. Conclusion: This study establishes the state of Sikkim as a new endemic area in India of Lyme disease besides its already reported endemicity in Arunachal Pradesh. No association was conclusively established between symptoms of Lyme and IgG seropositivity emphasizing the need for detailed history taking and clinical suspicion in endemic areas.

Comparative study of quality of leukoreduced packed red blood cell units as assessed by nageotte hemocytometry and flow cytometry.

PURPOSE: Assessment of residual white blood cell (rWBC) count is vital to ascertain the quality of leukodepleted (LD) blood components. Automated cell analyzers lack the sensitivity for the assessment of very few leukocytes as found in LD blood components. Flow Cytometry (FC) based methods and Nageotte hemocytometer are the most commonly used techniques for this purpose. The objective of this study was to compare the use of Nageotte hemocytometer and FC for quality control of LD red blood cell units. MATERIALS AND METHODS: A prospective, observational study was conducted in the Department of Immunohematology and Blood Transfusion of a tertiary care center from September 2018 to September 2020. About 303 LD-packed red blood cell units were tested by FC and Nageotte hemocytometer for rWBCs. RESULTS: The number of rWBC (mean) detected by flow cytometer and Nageotte's hemocytometer was 1.06 ± 0.43 white blood cell (WBC)/µL and 0.67 ± 0.39 WBC/µL, respectively. Coefficient of variation was 58.37% by Nageotte hemocytometer method and 40.46% by FC. Linear regression analysis did not show any correlation (R2= 0.098, P = 0.001) whereas Pearson's correlation coefficient showed a weak relation (r = 0.31) between the two methods. CONCLUSION: Flow cytometric technique provides a more precise and accurate objective tool compared to Nageotte hemocytometer which is labor intensive, time consuming, and prone to errors arising out of subjectivity along with reported underestimation bias. In the absence of adequate infrastructure, resources, and trained workforce, Nageotte hemocytometer method is a reliable alternative. Nageotte's chamber could be best used in the resource-constrained setup as it offers a relatively inexpensive, simple, and viable means to enumerate rWBCs.

Post vaccination antibody titres of hepatitis B surface antigen (anti-HBs) in a mixed cohort of health care workers.

Background: Hepatitis B (HepB) is an important vaccine preventable infection among healthcare workers (HCWs). Vaccination against Hep B virus, remains the foremost preventive approach. This study aims to measure the antibody titres to Hep B surface antigen (anti-HBs) in a mixed cohort of HCWs. It also aims to study the association between time since vaccination and the anti-HBs titres thus evaluating the duration of seroprotection. Methods: A total of 200 HCWs, including nursing students (n = 112), nursing staff (n = 49), laboratory technicians (n = 30) and doctors (n = 9) who had received all three doses of the Hep B vaccine and met the inclusion criteria of having taken all three doses of vaccine were included in this study. Anti-HBs titres were estimated by bioMérieux mini VIDAS® automated immunoassay based on the principle of enzyme-linked fluorescence assay. Results: Two hundred subjects aged 19 to 52 years were included in the study; mean age was 27.29 ± 0.568 years. Duration since vaccination in the study cohort was ≤ 5 years in 149 (74.5.0%), 6-10 years in 20 (10.0%) and >10 years in 31 (15.5%) subjects. Postvaccination antibody titres were > 100 mIU/ml in 85.0%, 10-100 mIU/ml in 11.0% and ≤ 10 mIU/ml in 3.5%. There was a decline noted in antibody titres as duration after vaccination increased. Increasing age was associated with falling protective titres. Conclusion: The study revealed that majority of the HCWs had adequate anti-HBs titres and were protected after vaccination.

Epidemiological analysis of SARS-COV-2 B.1.617.2 (delta variant) transmission in an educational institute.

Background: Despite having an effective COVID-19 vaccine, the COVID-19 pandemic is far from over and the delta variant continues to cause havoc across several continents. The present study was conducted to analyze and describe the occurrence of COVID-19 cases among completely vaccinated individuals. Methods: In an educational institute in Western Maharashtra, we analyzed a cluster of RTPCR positive COVID-19 cases among fully vaccinated students which occurred in 12 days. The cases were linked to a series of curricular and co-curricular events in the institute. A detailed epidemiological investigation and genome sequencing of cases were conducted. IgG antibodies against S1 protein of novel SARS-CoV-2 were estimated for cases and age, sex, and vaccination status matched controls. Results: All 37 identified cases were mild COVID. 188 high risk (HR) contacts of the cases were identified. The overall secondary attack was 9.5%. Out of 31 cases and 50 controls, 09 (29%) cases and 08 (16%) controls were found to have IgG antibodies against S1 protein of novel SARS-CoV-2 titer of more than 60 U/ml. Whole-genome sequencing of 15 samples of the cluster showed the presence of the Delta variant of SARS-CoV-2. No correlation was observed between Ct value and IgG S1 antibody titers. Conclusion: The study provides significant evidence that only vaccination alone does not completely protect against SARS-CoV-2 B.1.617.2 (Delta) variant infection. An all-encompassing multicomponent strategy involving implementation of NPIs, robust contact tracing, early identification and isolation of cases, and high vaccination coverage is the way forward for the prevention of COVID-19.

Effectiveness of ChAdOx1 nCOV-19 Vaccine: Experience of a tertiary care institute.

BACKGROUND: The first dose of the ChAdOx1 nCoV- 19 Corona Virus Vaccine (Covishield) was administered to the eligible beneficiaries of tertiary care institute of Western Maharashtra on 16 Jan 21 and in the past three months almost 97% of the staff has been vaccinated. The present study analyses the incidence of COVID cases in the unvaccinated and vaccinated population of the institute. METHODS: All Covid 19 infections (RT-PCR positive) from 01 February 21 to 25 April 21 were included in the study and analyzed as per their vaccination status. To assess the COVID 19 transmission in contacts, Secondary Attack Rates (SAR) of the pre-vaccination period (Jun-Oct 20) was compared with the present SAR. RESULTS: A total of 113 cases occurred in the study period (01 Feb to 25 Apr 21). Lower number of infections were observed among the fully vaccinated as compared to partially vaccinated and non-vaccinated. The overall vaccine effectiveness was found to be 88.6% (81.55-92.37) and 44.1% (4.55-67.3) in completely and partially vaccinated individuals respectively. Hazard Ratios for getting infected dropped significantly after 28 days of the second dose. The SAR in high risk contacts (HRCs) was found to be 4.25%, which was lower than SAR (20.6%) of pre-vaccination period. CONCLUSION: This is one of the earliest studies in India to report the impact of COVID-19 vaccination. The results indicate that the vaccine provides effective protection against COVID-19 infection. However, given the complex dynamics of vaccination, the role of NPIs and implementation of COVID appropriate behavior cannot be undermined.

Probable vertical transmission of severe acute respiratory syndrome coronavirus 2 infection from mother to neonate.

The pandemic spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has affected 188 countries and territories. Altered physiological status during pregnancy makes a mother vulnerable to severe SARS-CoV-2 infection. The virus may be transmitted from mother to baby during antenatal period or postnatal period. Although the primary mode of transmission of the virus is by respiratory droplets, there is emerging evidence of in utero transmission from mother to foetus. In this rare case report, we describe one such episode of probable vertical transmission. To the best of our knowledge, this is the second systematically investigated Indian case, indicating in utero transmission of SARS-CoV-2 from mother to foetus.

Whole genome sequence of colistin-resistant Escherichia coli from western India.

BACKGROUND: With virtually dried out new antibiotic discovery pipeline, emergence and spread of antimicrobial resistance is a cause for global concern. Colistin, a cyclic polypeptide antibiotic, often regarded as last resort for multi drug resistance gram-negative bacteria, is also rendered ineffective by horizontal transfer of resistance genes. Surveillance of colistin resistance in GNB is essential to ascertain molecular epidemiology. METHODS: Whole genome sequencing (WGS) of an unusual colistin resistant urinary isolate of Escherichia coli was performed using Illumina MiSeq platform using 2x250bp V2 chemistry by following the manufactures protocol (Illumina Inc. USA). Multiple web-based bio-informatic tools were utilized to ascertain antibiotic resistant genes. RESULTS: An approximate 5.4 Mb of genome of the urinary isolate AFMC_UC19 was sequenced successfully. Mobile colistin resistance gene (mcr) on the plasmid responsible for horizontal spread was absent in the isolate. CONCLUSION: Colistin resistance has been reported previously in Klebsiella pneumoniae and it is a rare occurrence in Escherichia coli in Indian setting. Although the isolate lack mcr mediated colistin resistance, emergence and spread of colistin resistant in gram-negative bacteria pose a threat.

Genotypic characterization of carbapenem resistant Enterobacterales in clinical isolates from western Maharashtra.

PURPOSE: To study distribution of carbapenemase genes namely; New Delhi metallo-beta-lactamase (blaNDM), Oxacillinase-48 (blaOXA48), Verona Integron-Encoded Metallo-beta-lactamase (blaVIM) and Imipenemase (blaIMP) in carbapenem resistant Enterobacterales (CRE), isolated from clinical samples. METHOD: This cross-sectional study was conducted at a tertiary care hospital of western Maharashtra over six months period. CREs were identified by conventional disc diffusion and modified carbapenem inactivation method (mCIM). A total of 50 consecutive CRE isolates from clinical samples were subjected to home brewed polymerase chain reaction (PCR) for detection of carbapenemases. RESULTS: Out of the 50 CRE isolates, at least one of the four carbapenemase genes was detected in 49 (98%) isolates. The frequency of distribution of these genes were NDM 90% (n = 45), OXA48 60% (n = 30) and VIM 12% (n = 6). Dual combination of blaNDM and blaOXA48 (50%) was the commonest pattern observed, which was frequently associated with Klebsiella pneumoniae. CONCLUSIONS: The study indicate high prevalence of NDM warranting strict anti-microbial stewardship practices. Surveillance of CRE and resistance mechanism is essential to monitor the trend and take informed decision for appropriate anti-microbial therapy.

Cost-effectiveness of a novel strategy of HIV/AIDS care in Armed Forces: A stochastic model with Monte Carlo simulation.

BACKGROUND: Despite the initial success of HIV/AIDS policy, an increasing number of patients are failing the first-line antiretroviral therapy (ART) each year and the failure rates are increasing. There is a need for identification of novel strategies to reduce failure rates. The aims of the study are (1) to design a novel strategy to reduce ART failure rates and (2) to create a stochastic model using Monte Carlo (MC) simulation comparing the novel strategy with existing strategy. METHODS: A novel strategy based on annual plasma viral load testing and resistance testing for HIV treatment at baseline and at the time of failure was designed. A cohort of 1000 patients each was created for the existing strategy and a novel strategy. Assumptions were included from Indian studies and own data. The two strategies were compared over 20 years of follow-up using stochastic modeling and MC simulation was done for death rates, failure rates, and cost-effectiveness analysis. SimVoi add-in software for MS Excel was used for simulations. Student's t-tests were performed for comparing continuous variables, and the cumulative rates for various outcomes were plotted using Kaplan-Meier analysis. RESULTS: The novel strategy resulted in lower mortality over a 20-year period (279.9 + 7.13 deaths vs 130.43 + 6.03 deaths) with incremental cost per life saved at Rs 32,925 per year. Incremental cost-effectiveness ratio cost per quality-adjusted life year was Rs 1.33 lakhs/annum at constant rate of discounting and just under Rs 90,000 per annum using differential discounting. CONCLUSION: Armed Forces are likely to benefit by adopting the novel strategy that is cost-effective with a significant mortality benefit.

Absence of Integrase Strand Transfer Inhibitor Associated Resistance in Antiretroviral Therapy Naïve and Experienced Individuals from Western India.

The Indian national AIDS control program heavily relies on low cost nucleoside reverse transcriptase inhibitor (NRTI) and non-nucleoside reverse transcriptase inhibitor (NNRTI). With global increase in resistance to these, alternative antiretroviral combinations need to be explored. Owing to higher potency, better efficacy and tolerability, recently WHO recommended integrase strand transfer inhibitor (INSTI) based first-line antiretroviral therapy (ART). There is lack of INSTI resistance surveillance data from India. Thus, there is a need to analyze integrase (IN) gene from primarily HIV-1 subtype C infected Indian population, before widespread introduction of INSTI in first-line ART. Plasma samples were collected from INSTI naïve individuals reporting to ART centre of Pune, India. RNA was extracted and IN gene was amplified by nested polymerase chain reaction (PCR) using prior published primers. PCR product of 867 bp was bi-directionally sequenced and resistance associated mutation were analyzed using Stanford University HIV drug resistance algorithm. A total of 58 HIV-1 sequences from 62 INSTI naïve individuals were successfully genotyped. Of these 58, 40 were ART naïve, newly diagnosed and remaining individuals were on NRTI, NNRTI, or protease inhibitors based failing regimen. The commonest subtype identified in the study was C (93%) followed by A1 (3.5%). A total of 191 (66.31%) fully conserved amino acid (aa) positions were observed in IN gene. Overall there was absence of major INSTI resistance mutation, however, E157Q (13.79%) emerged as common polymorphic mutation. Other accessory mutations were L74IM (34.48%), Q95K (1.72%), and T97A (1.72%). To conclude, this first Indian study on primarily HIV-1 subtype C sequences characterized aa variations in IN gene and indicated absence of major INSTI resistance associated mutations.

Measles, mumps, and rubella: A cross-sectional study of susceptibility to vaccine-preventable diseases among young people in India.

BACKGROUND: Global elimination of vaccine preventable diseases, such as measles, mumps and rubella is a priority. Many countries have reported diminishing of antibody titres against these diseases among young population as immunization coverage of adolescents and adults in not monitored. The objective of this study was to determine the susceptibility against measles, mumps and rubella among young adults. METHODS: In this cross-sectional study serological evidence of susceptibility to measles, mumps and rubella was determined by qualitative detection of IgG antibody titres by commercially available enzyme linked florescence assay (VIDAS, bioMerieux) in serum samples young adults. RESULTS: A total of 335 young individuals (mean age: 20.54 ± 1.37 years) participated voluntarily between May 2017 to September 2018, of which 183 (54.63%) were males. Seroprotection against measles, mumps and rubella were 87.16%, 82.69% and 79.10% respectively. CONCLUSION: Serological surveillance is important to monitor immune status in population. Susceptibility of young adults to measles, mumps, and rubella indicates need for booster vaccination. With the recent launch of measles-rubella vaccination campaign in India, country specific data will be required to plan periodicity of such campaign, which in turn would be based on accumulation of susceptible individuals in a community. Lastly, inclusion of mumps vaccine in the national universal immunization program needs consideration.

HIV drug resistance following a decade of the free antiretroviral therapy programme in India: A review.

OBJECTIVE: The objective of this review was to assess the burden of HIV drug resistance mutations (DRM) in Indian adults exposed to first-line antiretroviral therapy (ART) as per national guidelines. METHODS: An advanced search of the published literature on HIV drug resistance in India was performed in the PubMed and Scopus databases. Data pertaining to age, sex, CD4 count, viral load, and prevalence of nucleoside reverse transcriptase inhibitor (NRTI)/non-nucleoside reverse transcriptase inhibitor (NNRTI) DRM were extracted from each publication. Year-wise Indian HIV-1 reverse transcriptase (RT) sequences were retrieved from the Los Alamos HIV database and mutation analyses were performed. A time trend analysis of the proportion of sequences showing NRTI resistance mutations among individuals exposed to first-line ART was conducted. RESULTS: Overall, 23 studies (1046 unique RT sequences) were identified indicating a prevalence of drug resistance to NRTI and NNRTI. The proportion of RT sequences with any DRM, any NRTI DRM, and any NNRTI DRM was 78.39%, 68.83%, and 73.13%, respectively. The temporal trend analysis of individual DRM from sequences retrieved during 2004-2014 indicated a rising trend in K65R mutations (p=0.013). CONCLUSIONS: Although the overall burden of resistance against first-line ART agents remained steady over the study decade, periodic monitoring is essential. There is the need to develop an HIV-1 subtype C-specific resistance database in India.

Antiretroviral resistance following immunological monitoring in a resource-limited setting of western India: A cross-sectional study.

BACKGROUND: The free antiretroviral therapy (ART) program in India still relies on the clinico-immunological monitoring for diagnosis of treatment failure. As the nucleoside reverse transcriptase inhibitor (NRTI) backbone is shared in first- and second-line regimens, accumulation of drug resistant mutations (DRMs) can compromise the efficacy of NRTI. This study was undertaken to describe the pattern of HIV DRMs following immunological monitoring and investigate its impact on the cycling of NRTI between first- and second-line ART. METHODS AND FINDINGS: This cross-sectional study was performed at a state-sponsored ART clinic of Pune city in western India between January and June 2016. Consecutive adults receiving first-line ART with immunological failure (IF) were recruited for plasma viral load (PVL) estimation. Randomly selected 80 participants with PVL >1000 copies/mL underwent HIV drug resistance genotyping. Of these, 75 plasma sample were successfully genotyped. The median CD4 count and duration of ART at the time of failure were 98 (IQR: 61.60-153.50) cells/µL and 4.62 (IQR: 3.17-6.15) years, respectively. The prevalence of NRTI, non-NRTI, and major protease inhibitor resistance mutations were 89.30%, 96%, and 1.33%, respectively. Following first-line failure, sequences from 56.67% of individuals indicated low- to high-level resistance to all available NRTI. The proportion of sequences with ≥2 thymidine analogue mutations (TAMs) and ≥3 TAMs were 62.12% and 39.39%, respectively. An average of 1.98 TAMs per sequence were observed following IF as compared to 0.37 TAMs per sequence following targeted PVL monitoring at 12 months of ART from a prior study; this difference was significant (p<0.001). CONCLUSION: The option of cycling of NRTI analogues between first- and second-line regimens would no longer be effective if individuals are followed-up by immunological monitoring due to accumulation of mutations. Introduction of routine PVL monitoring is a priority for the long-term sustainability of free ART program in India.

Chryseobacterium Indologenes Pneumonitis in an Infant: A Case Report.

Chryseobacterium indologenes, a non-fermentative Gram-negative bacilli distributed widely in nature, is an emerging nosocomial pathogen, inherently resistant to a wide range of antibiotics. There is limited number of C. indologenes infections reported from India. We report a case of C. indologenes associated pneumonia in a three-month-old infant with congenital heart disease. This case highlights the importance of prompt diagnostic workup and targeted antibiotic therapy for its effective management.

Cross-sectional study of virological failure and multinucleoside reverse transcriptase inhibitor resistance at 12 months of antiretroviral therapy in Western India.

The free antiretroviral therapy (ART) program in India has scaled up to register second largest number of people living with HIV/AIDS across the globe. To assess the effectiveness of current first-line regimen we estimated virological suppression on completion of 1 year of ART. The study describes the correlates of virological failure (VF) and multinucleoside reverse transcriptase inhibitor (NRTI) drug resistance mutations (DRMs).In this cross-sectional study conducted between June and August 2014, consecutive adults from 4 State sponsored ART clinics of western India were recruited for plasma viral load screening at 12 ±â€Š2 months of ART initiation. Individuals with plasma viral load >1000 copies/mL were selected for HIV drug resistance (HIVDR) genotyping. Logistic regression analyses were performed to assess factors associated with VF and multi-NRTI resistance mutations. Criteria adopted for multi-NRTI resistance mutation were either presence of K65R or 3 or more thymidine analog mutations (TAMs) or presence of M184V along with 2 TAMs.Of the 844 study participants, virological suppression at 1 year was achieved in 87.7% of individuals. Factors significantly associated with VF (P < 0.005) were 12 months CD4 count of ≤100 cells/µL (adjusted OR -7.11), low reported adherence (adjusted OR -4.44), and those living without any partner (adjusted OR -1.98). In patients with VF, the prevalence of non-nucleoside reverse transcriptase inhibitor (NNRTI) DRM (78.75%) were higher as compared to NRTI (58.75%). Multi-NRTI DRMs were present in 32.5% of sequences and were significantly associated with CD4 count of ≤100 cells/µL at baseline (adjusted OR -13.00) and TDF-based failing regimen (adjusted OR -20.43). Additionally, low reported adherence was negatively associated with multi-NRTI resistance (adjusted OR -0.11, P = 0.015). K65R mutation was significantly associated with tenofovir (TDF)-based failing regimen (P < 0.001).The study supports early linkage of HIV-infected individuals to the program for ART initiation, adherence improvement, and introduction of viral load monitoring. With recent introduction of TDF-based regimen, the emergence of K65R needs to be monitored closely among HIV-1 subtype C-infected Indian population.

Short Communication: Limited HIV Pretreatment Drug Resistance Among Adults Attending Free Antiretroviral Therapy Clinic of Pune, India.

In India, the roll out of the free antiretroviral therapy (ART) program completed a decade of its initiation in 2014. The success of first-line ART is influenced by prevalence of HIV pretreatment drug resistance (PDR) in the population. In this cross-sectional study, we sought to determine the prevalence of PDR among adults attending the state-sponsored free ART clinic in Pune in western India. Fifty-two individuals eligible for ART as per national guidelines with median CD4 cell count of 253 cells/mm(3) (inter quartile range: 149-326) were recruited between January 2014 and April 2015. Population-based sequencing of partial pol gene sequences from plasma specimen revealed predominant HIV-1 subtype C infection (96.15%) and presence of single-drug resistance mutations against non-nucleoside reverse transcriptase inhibitor in two sequences. The study supports the need for periodic surveillance, when offering PDR testing at individual level is not feasible.

Near Full-Length Genomic Characterization of a Novel CRF 01_AE/C Recombinant from Western India.

HIV is known for its genetic variability across the globe. The HIV epidemic in India is primarily driven by subtype C, although sporadic circulating and unique recombinant forms are also reported from a few metropolitan cities in which genotyping facilities are available. Here we report a novel CRF01_AE/C recombinant from a multicenter study on the effectiveness of antiretroviral therapy (ART), 12 months after its initiation. Our subject is a 32-year-old heterosexual female, a native of Pune city in western India. Identification and analyses of recombination breakpoints using jpHMM@Gobics and SimPlot bootscanning revealed six recombination breakpoints, indicating insertion of the CRF01_AE genome at three points in the backbone of subtype C. Both subtype C and CRF01_AE are commonly seen in the population at risk of heterosexual HIV transmission, thereby providing an opportunity for cocirculation and recombination. The emergence of a novel recombinant of CRF01_AE/C is indicative of the increasing genetic diversity of the HIV epidemic in India.

Differentiation of wild-type varicella-zoster strains from India and the Oka vaccine strain using a VZV open reading frame - 62 based PCR-RFLP technique.

Since the introduction of varicella vaccination in India, surveillance of circulating VZV strains has gained significance. Differentiating wild-type VZV strains from the Oka vaccine strain can be achieved only by molecular genotyping methods. The development of PCR methods for VZV strain differentiation has been hampered by the fact that the VZV genome is highly conserved. We used VZV ORF 62 PCR-RFLP analysis to identify and differentiate wild-type VZV strains in India from the Oka vaccine strain. Digestion of VZV ORF 62 amplicons with SmaI, enabled accurate strain differentiation; the Oka strain was positive for three SmaI sites, compared to two SmaI sites in the wild-type VZV strains that we tested.

Differentiation of wild-type varicella-zoster strains from India and the Oka vaccine strain using a VZV open reading frame - 62 based PCR-RFLP technique

Since the introduction of varicella vaccination in India, surveillance of circulating VZV strains has gained significance. Differentiating wild-type VZV strains from the Oka vaccine strain can be achieved only by molecular genotyping methods. The development of PCR methods for VZV strain differentiation has been hampered by the fact that the VZV genome is highly conserved. We used VZV ORF 62 PCR-RFLP analysis to identify and differentiate wild-type VZV strains in India from the Oka vaccine strain. Digestion of VZV ORF 62 amplicons with SmaI, enabled accurate strain differentiation; the Oka strain was positive for three SmaI sites, compared to two SmaI sites in the wild-type VZV strains that we tested.