Outbreak of cholera in a remote village in western India.

Background & objectives: Atypical El Tor strains of Vibrio cholerae are frequently implicated in outbreaks of cholera. It is important to understand genetic variations of such strains which impact clinical and epidemiological outcomes. The present study was carried out to characterize an outbreak of cholera which occurred between July 8 and 13, 2018, in a remote settlement in Nashik district, Maharashtra. Methods: A large number of acute diarrhoea cases were reported in Rahude village, Nashik, Maharashtra since July 8, 2018. Molecular characterization of the isolated strains of V. cholerae was done. Results: 195 cases of cholera were detected from a population of 850 (attack rate 22.9%) with two deaths (Case Fatality Ratio of 1.03). A non-haemolytic polymyxin B sensitive strain of V. cholerae O1 Ogawa was isolated from 5/14 fecal samples. Molecular characterization of the isolates indicated that this strain was an altered El Tor (AET) strain. Deletion of the trinucleotide 'GTA' in the rstB gene, a unique feature of classical strains, was observed. Interpretation & conclusions: A cholera outbreak caused by a non-haemolytic polymixin B sensitive AET strain, occurred from July 8 to 13, 2018, in a remote settlement in western India. The molecular characterization of the outbreak strains highlighted an assortment of genetic determinants, stressing the need to monitor the genetic attributes of V. cholerae O1 in outbreaks for better understanding and mapping of clinical and epidemiological changes.

A Multicenter, Randomized, Open-Label Trial Comparing the Efficacy and Safety of Monoclonal Anti-Rh (D) Immunoglobulin with Polyclonal Anti-Rh (D) Immunoglobulin for the Prevention of Maternal Rh-Isoimmunization.

OBJECTIVES: To compare the efficacy and safety of monoclonal anti-Rhesus (anti-D) immunoglobulin (IgG) with polyclonal anti-D IgG in the prevention of maternal Rh-isoimmunization. METHODS: This was a randomized, multicenter, open-label, comparative clinical trial conducted in the obstetric in-patient departments of nine tertiary care hospitals in India. 206 Rhesus (D)-negative women, not sensitized to Rh antigen, and delivering Rh positive babies, received postpartum intramuscular administration of monoclonal or polyclonal anti-D IgG. The main outcome measures were the proportion of subjects protected from Rh-isoimmunization, identified by a negative indirect Coombs test (ICT) result, at day 180 after anti-D IgG administration, and incidence of adverse events. RESULTS: 105 subjects were randomized to the monoclonal group and 101 to the polyclonal group. 94 from the monoclonal group had a negative ICT result and none had a positive ICT result at day 180, whereas 87 from the polyclonal group had a negative ICT result and one had a positive ICT result; the rest (11 and 13 subjects respectively) were lost to follow-up. A total of 5 adverse events were reported (3 in the monoclonal group and 2 in the polyclonal group); only one of these was serious. All the adverse events were judged to be unrelated to the interventional drug. None of the subjects in the monoclonal group developed immunogenic reaction to the monoclonal anti-D. CONCLUSION: The efficacy and safety of the monoclonal preparation of anti-D was comparable to the polyclonal preparation of anti-D when used in the prevention of maternal Rh-isoimmunization.Trial registration Clinical Trial Registration Number: CTRI/2015/09/006172.

Molecular characterization of group A rotavirus (RVA) strains detected in bovine and porcine species: Circulation of unusual rotavirus strains. A study from western, India.

Group A rotaviruses (RVA) are considered as important causative agents of diarrhea in both human and animal species. Fecal specimens (n = 300) were collected from both diarrheic and healthy animals during the year 2009 from animal farms from Nagpur (Maharashtra), Western India. RVA antigen was detected by ELISA in 3.1-25% and 72% in bovine and porcine species, respectively. Genotyping based on VP6, VP7 and VP4 of RVA-positive samples showed predominance of genotype I-1 (63%) and genotype I-2 (37%), G4 (45.5%) and G10 (27.3%) genotypes, P[6] (72.7%) and P[8] (18.1%) genotypes, respectively. Other RV genotypes such as G1(4.5%), G2(9.1%), G3(4.5%) and mixed infections (9.1%) were detected at low level. Predominance of unusual G-P combinations (9/23, 39.1%) were observed. Circulation of G2P[8] and mixed infections with G1, G3, P[6] and G1, P[8], P[6]) are reported in porcine species for the first time in Western India. In conclusion the present study highlights the circulation of unusual G-P combinations and VP6 genogroup specificities of human RVA strains indicative of possible interspecies transmission and reassortment events in animal species. The study further warrants utmost need for such surveillance studies across the country to understand the role of animals as genetic reservoirs for the emergence of RVA strains pathogenic for humans. Keywords: rotaviruses; genotypes; unusual G-P types; animals.

Epidemiology and genetic diversity of human parechoviruses circulating among children hospitalised with acute gastroenteritis in Pune, Western India: a 5-years study.

Human parechoviruses (HPeVs) are known to cause various clinical manifestations including acute gastroenteritis. Although HPeV infections and their genotypes have been detected in human patients worldwide, no such reports are available from India to ascertain the association of HPeVs in acute gastroenteritis. The present study was conducted to determine the clinical features and genetic diversity of HPeVs detected in children hospitalised for acute gastroenteritis. Stool specimens (n = 979) collected from children aged ⩽5 years hospitalised for acute gastroenteritis in Pune, western India during January 2006-December 2010 were included. HPeV RNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) (5'UTR) followed by genotyping using VP1 gene-based PCR and phylogenetic analysis. HPeV was detected in 13·9% (136/979) of the cases, co-infections with other enteric viruses were found in 43·4%. HPeV was more frequent in children ⩽1 year age with infections reported throughout the year. A total of 102/136 (75%) HPeV strains were genotyped, which comprised 13 different HPeV genotypes. Of these, HPeV1 was the most predominant genotype detected and phylogenetically clustered with the Harris strain which is rarely reported. The study documents circulation of heterogeneous HPeV genotypes. Two variant strains of HPeV4 and 'RGD absent' HPeV5 and 6 strains were also detected. This is the first report of HPeV with diversified genotypes identified in acute gastroenteritis patients from India.

Epidemiological profile and genetic diversity of sapoviruses (SaVs) identified in children suffering from acute gastroenteritis in Pune, Maharashtra, Western India, 2007-2011.

Sapoviruses (SaVs) are responsible for sporadic cases and outbreaks of acute gastroenteritis. Despite this, few studies in India have focused on the epidemiological investigation of SaV in cases of acute gastroenteritis. The aim of this study was to understand the molecular epidemiology, genetic diversity and clinical impact of SaV in diarrhoeic children from Pune, Western India. Between 2007 and 2011, a total of 985 faecal samples from diarrhoeic cases and non-diarrhoeic controls were collected and examined for the presence of SaV by nested RT-PCR. SaV was detected in 2·7% (21/778) of the cases and 1·9% (4/207) of the controls. We observed that the majority of SaV mono-infections caused severe gastroenteritis (67%) with clinical manifestations of diarrhoea (100%), vomiting (73%) and dehydration (80%). All known human SaV genogroups were detected in the study. At least eight genotypes were identified from cases and controls. Genogroups GIV and GV, along with genotypes GI.5, GII.4 and GII.6, were discovered for the first time in India. Two GII.4 study strains were found to be 98·5-99% identical, having a novel intra-genogroup recombinant (GII.1/GII.4) recently reported from the Philippines, suggesting probable evidence of recombination. The circulation pattern of SaV genotypes varied during the study period, with GII.1 being predominant in 2007 and 2009, GIV.1 in 2008, and GV.1 in 2011.

Group C rotavirus infection in patients with acute gastroenteritis in outbreaks in western India between 2006 and 2014.

Faecal specimens collected from outbreak (n = 253) and sporadic (n = 147) cases of acute gastroenteritis that occurred in western India between 2006 and 2014 were tested for group C rotavirus (GCR) using partial VP6 gene-based RT-PCR. All specimens were tested previously for the presence of other viral and bacterial aetiological agents by conventional methods. The rate of GCR detection was 8·6% and 0·7% in outbreak and sporadic cases, respectively. GCR infections prevailed in outbreaks reported from rural areas (10·9%) compared to urban areas (1·6%). Clinical severity score of the patients with GCR infection (n = 23) indicated severe disease in the majority (70%) of cases. The age distribution analysis indicated 52·1% of GCR infections in children aged <10 years. The male:female ratio in GCR-positive patients was 2·3:1. Of the 23 GCR-positive cases, 17 (73·9%) had a sole GCR infection and six had mixed infections with other viral and/or bacterial agents. Phylogenetic analysis of nucleotide sequences classified GCR strains of the study in to I2 genotype of the VP6 gene. This is the first study to show the occurrence of GCR in gastroenteritis outbreaks in India.

Clinico-epidemiology and genetic diversity of Salivirus in acute gastroenteritis cases from Pune, Western India: 2007-2011.

Acute gastroenteritis is a leading cause of mortality in children from developing countries. Recently, Salivirus has been frequently detected in acute gastroenteritis patients, suggesting its possible aetiological role. Conflicting reports available on disease association of Salivirus have made it difficult to ascertain their causative role. The overall epidemiology and clinical features of Salivirus infections are poorly understood. The present five year study was undertaken to investigate the presence and genetic diversity of Salivirus in acute gastroenteritis cases from Pune, Western India and to determine the clinico-epidemiological features of Salivirus infections. A total of 985 faecal samples (778 acute gastroenteritis and 207 asymptomatic controls), collected from three local hospitals (Jan2007-Dec2011) were examined for the presence of Salivirus by RT-PCR. Molecular characterization was performed by PCR amplification of the 3D and VP regions. Frequency of Salivirus detection in cases (2.6%) and controls (1.93%) was not significantly different (p = 0.57). Co-infection with other enteric viruses was seen in 50% of the cases. Comparison of clinical features between Salivirus mono and mixed infections revealed that Salivirus alone did not exacerbate gastroenteritis. The frequency of diarrhoea and overall clinical severity of mixed infections was significantly greater than mono infections (p = 0.02). Based on clinical findings, our study suggests that Salivirus does not cause severe gastroenteritis. Phylogenetic analysis indicated that study strains belonged to Salivirus A1 and formed 2 distinct clusters which shared nucleotide identities of 94.1-96.2% and 88.9-93.8% between themselves in 3D and VP regions, respectively. Interestingly, the more divergent Cluster2 strains shared a low nucleotide identity with the closest reference strain in both regions (~95% in 3D and ~92% in VP) suggesting that they could represent a variant type of Salivirus A1. The genetic diversity in strains detected from study region, emphasizes the need for Salivirus surveillance from other regions of India.

Molecular surveillance of non-polio enterovirus infections in patients with acute gastroenteritis in Western India: 2004-2009.

Acute gastroenteritis is a major cause of childhood morbidity and mortality worldwide. Rotavirus (RV) and Norovirus (NoV) are the leading cause of the disease. Despite the use of improved diagnostic methods a significant proportion of gastroenteritis cases remained undiagnosed. Though nonpolio enteroviruses (NPEVs) have been reported frequently in children with acute gastroenteritis, their etiologic role has not been established. To investigate the epidemiology of NPEVs in gastroenteritis cases which remained negative for leading causative agents, 955 RV and NoV negative stool specimens from children hospitalized for acute gastroenteritis were included in the study. A case control study was conducted which includes stool specimens from 450 children with gastroenteritis and 162 asymptomatic control subjects to determine the association of NPEVs with the disease. NPEV detection and typing was carried out by RT-PCR and sequencing. Presence of RV, NoV, Adenovirus, and Astrovirus was confirmed by ELISA or PCR/RT-PCR. Overall 14% NPEV prevalence was noted. The percentage of children with NPEV infection differed significantly between gastroenteritis and non-gastroenteritis patients (13.7% vs. 4.9%). NPEV was more prevalent among patients with gastroenteritis of undetectable etiology as compared to those detected positive for other viruses (17.9% vs. 7%) (P < 0.01). Genotyping of NPEV identified predominance of EV-B species (56.5%) followed by EV-C (16.7%), EV-A (13.8%) species and mixed NPEV infections (13%). These data support the association of NPEVs with acute gastroenteritis and highlights the clinical and epidemiological features of NPEV infections in patients with acute gastroenteritis from western India.

Circulation of multiple enterovirus serotypes causing hand, foot and mouth disease in India.

Hand, foot and mouth disease (HFMD), a common contagious disease that usually affects children, can be caused by enteroviruses. Coxsackievirus A16 (CV-A16) and enterovirus 71(EV-71) are the major aetiological agents of HFMD. Other EV serotypes, CV-A4-7, CV-A9-10, CV-B1-3, CV-B5, E-4 and E-19, have also been found associated with both sporadic infections and outbreaks of HFMD. In India, outbreaks of HFMD have been documented; however, molecular characterization of the aetiological agents has rarely been reported. Cases of HFMD were identified during 2009-2010 on the basis of clinical features in southern and eastern parts of India. The aim of the present study was to detect and characterize the aetiological agents associated with the disease. A total of 89 specimens consisting of 41 sera, 24 vesicular fluids, 18 stools and 6 throat swabs were collected from 61 clinically diagnosed HFMD cases from southern and eastern parts of India. RT-PCR followed by sequencing of PCR amplicons and phylogenetic analysis were performed on all specimens for detection of EV RNA and identification of EV types. EV RNA was detected in 47.1 % (42/89) of the specimens collected from 57.4 % (35/61) of the HFMD cases. Thirty-six of 42 EV strains showed amplification of the VP1/2A junction or VP1 regions. Sequence analysis of the amplicons identified the presence of CV-A16 (54.8 %), CV-A6 (38.1 %), EV-71 (2.4 %), CV-A10 (2.4 %) and E-9 (2.4 %) serotypes in the HFMD cases. The study documents CV-A16 and CV-A6 as major and CV-A10, EV-71 and E-9 as rare viral pathogens of HFMD in India.

Circulation of Aichi virus genotype B strains in children with acute gastroenteritis in India.

Acute gastroenteritis (AG) is considered as one of the major health problems affecting humans of all ages. A number of viruses have been recognized as important causes of this disease. Recently, Aichi virus has been shown to play an aetiological role in sporadic infections and outbreaks of AG. A study on surveillance of enteric viruses was conducted during 2004-2008 in three cities in Maharashtra state, western India. A total of 1240 stool specimens from children aged ≤8 years hospitalized for AG were screened for the presence of Aichi virus by RT-PCR of the 3C-3D junction region followed by sequencing for the identification of genotype. Aichi virus was detected at a prevalence of 1·1% in the <5 years age group and characterized as genotype B. This is the first report on the circulation of Aichi virus genotype B in India.

Astrovirus associated acute gastroenteritis in western India: predominance of dual serotype strains.

A five-year (2004-2008) study was conducted on patients with acute gastroenteritis from different cities of Maharashtra, western India to detect and characterize astrovirus infections. A total of 1340 fecal specimens were collected from sporadic cases that included 1240 children (

Enteroviruses in patients with acute encephalitis, uttar pradesh, India.

An outbreak of viral encephalitis occurred in northern India in 2006. Attempts to identify an etiologic agent in cerebrospinal fluid by using reverse transcription-PCR showed positivity to enterovirus (EV) in 66 (21.6%) of 306 patients. Sequencing and phylogenetic analyses of PCR products from 59 (89.3%) of 66 specimens showed similarity with EV-89 and EV-76 sequences.

Antiviral activity of the Indian medicinal plant extract Swertia chirata against herpes simplex viruses: a study by in-vitro and molecular approach.

PURPOSE: The antiviral activity of Indian Medicinal plant extract Swertia chirata was tested against Herpes simplex virus (HSV) type-1, using multiple approaches both at cellular and molecular level. METHODS: Cytotoxicity, plaque reduction, virus infectivity, antigen expression and polymerase chain reaction (PCR) assays were conducted to test the antiviral activity of the plant extract. RESULTS: Swertia plant crude extract (1 gm/mL) at 1:64 dilution inhibited HSV-1, plaque formation at more than 70% level. HSV antigen expression and time kinetics experiments conducted by indirect immunofluorescence (IFA) test, revealed a characteristic pattern of small foci of single fluorescent cells in Swertia extract treated HSV-1 infected cells at 4 hours post infection dose, suggested drug inhibited viral dissemination. Infected cell cultures treated with Swertia extract at various time intervals, tested by PCR, failed to show amplification at 12, 24-72 hours. HSV-1 infected cells treated with Acyclovir (antiviral drug) did not show any amplification by PCR. CONCLUSIONS: In this preliminary study, the Indian medicinal plant extract, Swertia chirata showed antiviral properties against Herpes simplex virus type-1.

Detection of Mycoplasma species in cell culture by PCR and RFLP based method: effect of BM-cyclin to cure infections.

PURPOSE: A two-stage nested polymerase chain reaction (PCR) assay system was described that amplifies the 16S-23S rRNA spacer region sequences of Mycoplasma and Acholeplasma infections in cell cultures and virus stocks. METHODS: Established cell lines and virus stocks were screened for the presence of Mycoplasma by using nested PCR using two sets of outer and inner primers, amplifies 16S-23S rRNA. PCR and restriction fragment length polymorphism (RFLP) assay was used to detect and identify most of the species-specific Mycoplasmas involved in cell cultures and virus stock contaminants. Infected cultures detected by PCR-RFLP were further treated with BM-cyclin (5 microg/mL) and passaged for three times and tested for Mycoplasma infections by PCR-RFLP. RESULTS: Mycoplasma pirum and Mycoplasma orale infections were detected by nested PCR. Species specificity was identified by using RFLP of Vsp I, Cla I and Hin dIII restriction enzymes. Mycoplasma infections were cured by treatment with BM-cyclin. This was further confirmed by non-amplification of PCR amplimers in BM-cyclin treated vs. non-treated cultures. CONCLUSIONS: Regular monitoring of cell cultures for Mycoplasma infections and identification of species-specific Mollicutes will identify the source of contaminations. This approach can be used for quality control of the biological reagents used in cell culture and virology laboratories.

Outbreak of acute hemorrhagic conjunctivitis in Maharashtra and Gujarat states of India, caused by Coxsackie virus A-24 variant.

Acute hemorrhagic conjunctivitis is associated with enteroviruses. Among these, Coxsackie A-24 variant (CA-24) and Enterovirus-70 (EV-70) are known to cause epidemics and pandemics. An outbreak of acute hemorrhagic conjunctivitis occurred in August-September 2003 in Maharashtra and Gujarat states of India. The present investigation was carried out to determine the viral etiological agent associated with the epidemic. Virus isolates were obtained from 11 eye swabs of conjunctivitis patients using HeLa/ Hep-2 cell lines. The isolates were characterized by serological and mouse pathogenecity tests, RT-PCR using enterovirus common primers (VP4-VP2), CA-24 specific primers (3C-proteinase region), EV-70 primers (VP-3) followed by sequencing, and phylogenetic analysis. The virus was characterized as a Coxsackie A-24 variant (CA-24v) and none of the isolates were found to be positive for EV-70. Sequencing of the PCR products derived from all the 11 isolates revealed 98.4% (SE 0.20) nucleotide identity within the Indian strains and 98.6% (0.50) and 94.4% (0.30) nucleotide identity respectively with the West Indies and Asian strains reported worldwide. The findings suggest that the outbreak of acute hemorrhagic conjunctivitis that occurred in Maharashtra and Gujarat states of India during August-September 2003 was caused by the Coxsackie A-24 variant (CA-24v).

Cervical squamous intra-epithelial changes and human papillomavirus infection in women infected with human immunodeficiency virus in Pune, India.

In view of the dual burden of HIV infection and cervical cancers in India, this study was undertaken to estimate the prevalence of Pap smear abnormalities and human papillomavirus infection among HIV-infected women. Consecutive HIV-infected women attending voluntary counseling testing clinics were enrolled. Written informed consent, demographic information, Pap smears, cervical swabs for HPV typing and a blood sample for CD4+ cell count were collected. Treatment for opportunistic and sexually transmitted infections and reproductive tract infections was provided. Women with Pap smear abnormality were referred for further intervention. Between January 2003 and May 2004, 287 HIV-infected women were enrolled. Pap smear abnormalities were seen in 6.3% women and were more common among women aged 30 and above (P=0.042) and those who had suffered from opportunistic infections (P=0.004). In multivariate analysis, Pap smear abnormalities were associated independently with opportunistic infections (P=0.02, AOR 3.8, 95% CI 1.2--11.5). Of the 100 random cervical specimens screened for HPV 16 and 18 genotypes, 33% (95 CI 23.9--43.1) were positive for HPV 16/18. Of the 122 patients who returned for a follow-up visit, 5 patients (4.1%) who did not have Pap smear abnormality at baseline, had developed Pap smear abnormality. The incidence of Pap smear abnormalities was 5.5 per 100 person year of follow-up. In order to prevent thousands of deaths due to cervical cancer in India, there is a need for strengthening the Pap smear screening program and HPV vaccine development.

Polymorphism of the p53 codon 72 Arg/Pro and the risk of HPV type 16/18-associated cervical and oral cancer in India.

Infection of high risk human papillomaviruses (HPVs) specifically the types 16 and 18 has been strongly implicated in the development of cervical cancer. The E6 oncoproteins of these high risk HPVs are known to bind and induce degradation of p53 tumour suppressor protein through the ubiquitin pathways. This degradation is controlled by a common polymorphism of the p53 gene encoding either a proline or an arginine at its codon 72 in exon 4. Recently, it has been demonstrated that the presence of homozygous arginine at codon 72 renders p53 about seven times more susceptible to E6-mediated proteolytic degradation as well as to cervical cancer than those with proline homozygotes or proline/arginine heterozygotes. In India, prevalence of HPV as well as cancers of the uterine cervix and the oral cavity are highest in the world. We have examined this allele-specific predisposition in cervical and oral cancer which is associated with HPV as well as in a non-HPV-linked cancer of the breast. We have carried out investigation in women comprising whole spectrum of cervical lesions with 128 HPV 16/18 positive and 35 HPV negative invasive cervical carcinomas and 34 cases of HPV (16/18) positive and 16 HPV negative cervical dysplasias (mild, moderate and severe) and 104 age-group-matched healthy women as controls. Additionally, we have analysed p53Arg-Pro polymorphism in 13 high risk HPV positive and 31 HPV negative oral cancers along with 20 normal controls and 77 breast cancers with 41 age-matched healthy controls. We observed more than two fold higher risk for homozygous arginine (chi2 = 6.3, df = 2, p = 0.04; OR = 2.3; 95% CI: 1.08-5.16) for HPV 16/18-positive cervical carcinomas when comparison was made only between HPV positive cervical cancers and normal controls but most interestingly, no significant association either in the frequency of homozygous arginine or proline alleles or their heterozygotes could be observed when all the three groups i.e. HPV-positive, HPV-negative cervical cancers and controls were considered simultaneously. No difference was also observed for either arginine or proline polymorphism between women with precancerous lesions of the uterine cervix carrying HPV 16/18 infection and controls. Similarly, increased risk of oral or breast cancer could not be correlated with the polymorphism of arginine/proline allele. Thus the interaction between HPV oncoproteins and the p53 gene polymorphism specifically, homozygous arginine at codon 72 appears to play no role in the development of either cervical or oral cancer and also it can not serve as a biomarker for early identification of cervical, oral or breast cancer.

A simple 'paper smear' method for dry collection, transport and storage of cervical cytological specimens for rapid screening of HPV infection by PCR.

Human papillomaviruses (HPVs) are major pathogens associated with the development of cancer of the uterine cervix, the most common malignant tumour of women worldwide. Reliable diagnosis of HPV infection, particularly the 'high-risk' types (16/18), may facilitate early identification of 'high-risk' populations for developing cervical cancer and may augment the sensitivity and specificity of primary cervical cancer screening programmes by complementing the conventional Pap test. A simple paper smear method has been developed for dry collection, transport and storage of cervical smears/scrapes at room temperature for subsequent detection of HPV DNA by PCR assay. Imprint biopsies, blood and fine-needle aspirates were also collected by this method. The cervical scrapes or other body fluids were smeared (within 0.5-1 cm diameter) and dried on to sterile small slides made of Whatman 3MM filter paper, and stored individually at room temperature or at 4 degrees C. A small piece (2-3 mm) of the paper smear was punched or cut out with a sterile surgical blade, boiled in an eppendorf tube containing 50 microl of distilled water for 5 min and used directly for PCR amplification. The quality and quantity of DNA derived from paper smears and the results of PCR amplifications for HPV type 16, BRCA1 and p53 genes were identical to those obtained from the same samples following collection in PBS, storage (-70 degrees C) and phenol-chloroform-based DNA extraction. DNA was stable in the paper smears for up to a year, whether stored at room temperature or at 4 degrees C. This method is simple, rapid and cost-effective, and can be effectively employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory.

Chlamydia trachomatis and human papillomavirus infection in Indian women with sexually transmitted diseases and cervical precancerous and cancerous lesions.

OBJECTIVES: Sexually transmitted diseases (STDs) and anogenital cancers are the major health problems in Indian women but no reliable estimate of the prevalence of either genital chlamydial infection or human papillomavirus (HPV) infection in STD patients is available. The aim of this study was to detect the frequency of Chlamydia trachomatis and the most prevalent high-risk HPV type 16 (HPV 16) infection in Indian women, with STDs and precancerous and cancerous lesions of the uterine cervix by polymerase chain reaction (PCR), and their comparison with those of conventional serology and antigen tests used for C. trachomatis detection. METHODS: Endocervical swabs or scrapes were collected from 50 women with STDs and 30 normal healthy women attending the STD clinics of Smt. Sucheta Kripalani Hospital, New Delhi. Scraped cervical cell specimens were also collected from 50 women with precancerous and cancerous lesions of the uterine cervix. Detection of C. trachomatis and HPV was carried out by PCR using chlamydia and HPV genome-specific oligonucleotide primers. The detection of chlamydial antigen and IgG-specific antibodies was carried out by enzyme immunoassay (EIA) and serological enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: A chlamydia plasmid-based PCR assay detected 50% (25 of 50) positivity of C. trachomatis in STD patients and HPV 16 DNA was found in 30% (15 of 50) of these cases which are significantly higher than those found in healthy controls. The PCR estimate of chlamydia was found to be higher than its reported frequency by tissue culture. The EIA could detect chlamydial antigen in only 13 cases (26%) while serological ELISA revealed evidence of chlamydia IgG-specific antibodies in 26 (52%) cases. Interestingly, in women with precancerous and cancerous lesions, the rate of HPV 16 infection was very high (52% and 72%, respectively), whereas the frequency of chlamydia infection was found to be 12-22% only. Occurrence of other sexually transmitted agents was also evaluated in the women. CONCLUSIONS: This is the first PCR estimate of genital chlamydial (50%) and HPV 16 (30%) infection in STD patients and women with precancerous and cancerous lesions of the uterine cervix in India. The PCR method seems to be a good alternative to tissue culture.