Arti Gulati

Can human papillomavirus DNA testing of self-collected vaginal samples compare with physician-collected cervical samples and cytology for cervical cancer screening in developing countries?

BACKGROUND To determine human papillomavirus (HPV) types by polymerase chain reaction (PCR)-reverse line blot assay and examine the concordance between HPV by Hybrid Capture 2 (HC2) and PCR on self-collected vaginal and physician-collected cervical samples and cytology. METHODS This was a cross-sectional study of 546 sexually active women aged > or =30 years with persistent vaginal discharge, intermenstrual or postcoital bleeding or an unhealthy cervix. Participants self-collected vaginal samples (HPV-S) and physicians collected cervical samples for conventional Pap smear and HPV DNA (HPV-P) testing and performed colposcopy, with directed biopsy, if indicated. HPV testing and genotyping was done by HC2 and PCR reverse line blot assay. Concordance between HC2 and PCR results of self- and physician-collected samples was determined using a Kappa statistic (kappa) and Chi-square test. RESULTS Complete data were available for 512 sets with 98% of women providing a satisfactory self-sample. PCR detected oncogenic HPV in 12.3% of self- and 13.0% of physician-collected samples. Overall, there was 93.8% agreement between physician-collected and self-samples (kappa=76.31%, 95% confidence interval [CI]: 64.97-82.29%, p=0.04)-complete concordance in 473 cases (57 positive, 416 negative), partial concordance in seven pairs and discordance in 32 pairs. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of self-sampling for detection of cervical intraepithelial neoplasia (CIN)2+ disease were 82.5%, 93.6%, 52.4% and 98.4%, respectively; for physician-sampling they were 87.5%, 93.2%, 52.2% and 98.9%, respectively; and for cytology they were 77.5%, 87.3%, 34.1% and 97.9%, respectively. Concordance between HC2 and PCR was 90.9% for self-samples (kappa=63.7%, 95% CI: 55.2-72.2%) and 95.3% for physician-collected samples (kappa=80.4%, 95% CI: 71.8-89.0%). CONCLUSIONS Self-HPV sampling compares favourably with physician-sampling and cytology. A rapid, affordable, HPV self-test kit can be used as the primary method of cervical cancer screening in low-resource situations.

Evaluation of cervical screening in rural North India.

To assess the accuracy of cervical screening with visual inspection and cytology testing, and the cure rate of cervical intraepithelial neoplasia (CIN) after treatment, in a rural population in North India.

Human papillomavirus-type distribution in women with and without cervical neoplasia in north India.

Our objective was to determine the human papillomavirus (HPV)-type prevalence in cervical samples in women with and without cervical neoplasia in an opportunistic hospital-based cancer-screening program. A cross-sectional study of 524 women presenting from January 2003 through June 2005 with symptoms of persistent vaginal discharge, intermenstrual bleeding, and postcoital bleeding or detected to have an unhealthy cervix underwent HPV genotyping by consensus polymerase chain reaction and reverse line-blot hybridization assay, conventional Pap smear, and colposcopy, with directed biopsy from all lesions detected. The prevalence rates of HPV infection among women with normal, low-grade cervical neoplasia (CIN 1) and high-grade CIN (>CIN2) were found to be 7.6%, 42.3%, and 87.5%, respectively. Seventeen high-risk and 6 low-risk HPV types were identified by the reverse line-blot assay. Multiple infections were seen in 20% of women. In normal women, the 6 commonest types were HPV-16, HPV-89, HPV-39, HPV-52, HPV-62, and HPV-18, whereas in high-grade disease, these were all high-risk types HPV-16, HPV-18, HPV-33, HPV-39, HPV-35, and HPV-56. HPV-16 was the commonest type in all groups, seen in 49.4% cases overall and in 74.3% of high-grade squamous intraepithelial lesion. It was followed by HPV-18 (7.4%) and HPV-33 and HPV-39 (4.9% each). HPV-89 was the commonest low-risk type (9.9%). HPV-16/18 were associated with 34.3% of normal, 45.4% of low-grade and 65.7% of high-grade lesions. A wide spectrum of HPV types is seen in north Indian women, with the majority being HPV-16 in all grades of histology. A vaccine against HPV-16 and HPV-18 could prevent two thirds of cases of high-grade cervical neoplasia.

Human papillomavirus type distribution in cervical cancer in Delhi, India

This hospital-based study in New Delhi, North India was performed to evaluate the prevalence of human papillomavirus (HPV) in cases of invasive cervical carcinoma (ICC). A total of 10 cases presenting with an obvious cervical growth were included in this study. 108 cases that was shown to be ICC on histology (101 squamous cell carcinomas, 4 adenocarcinomas, and one neuroendocrine carcinoma) were included in the analysis. DNA was extracted from tumor tissue and HPV genotype was determined by a consensus PCR assay using a reverse line blot hybridization assay. Of 106 evaluable cases, 104 (98.1%) were positive for HPV infection. Twelve different high-risk HPV types were found. There were 125 infections, 119 of which were high risk. Six cases had associated low risk infections. HPV 16 was the commonest type, seen in 73.6% cases followed by HPV 18 (14.2%) and 45 (11.3%). A vaccine with 100% efficacy in prevention of HPV 16 and 18 infections would theoretically reduce the total cancer burden in New Delhi by more than 75% (assuming 100% coverage). Increasing the genotype spectrum (e.g. valency) if the existing vaccines would be expected to have only a modest impact on the potential for cervical protection.

Prevalence of human papillomavirus infection among young women in North India

BACKGROUND The number of women infected with human papillomavirus (HPV) and the distribution of the HPV genotypes vary across populations and with age. OBJECTIVE To determine the prevalence and genotype distribution of HPV in young married women aged 16-24 years. METHODS 1300 women residing in an urban slum in Delhi donated samples of exfoliated cervical cells that were collected by the Digene((R)) kit and tested for the presence of HPV DNA by two techniques in parallel, i.e., PCR using PGMY consensus primers for all HPV types and the Digene HPV test (Hybrid Capture 2 (HC2) Probe B for high-risk (hr) types. Genotyping was done on all HPV positive samples using the Roche reverse line blot assay. RESULTS HPV infection was detected in 91/1300 (7%) samples by PCR and 110/1300 (8.4%) samples by HC2. Genotyping identified 20 high-risk and 11 low-risk types. HPV16 was the commonest high-risk type (3%) followed by HPV52 (1.2%) and HPV51 (0.8%). Among low-risk types, HPV62 was the commonest (0.8%), followed by HPV84 and HPV89 (0.5% each). Multiple infections were found in 3% of the HPV positive samples. CONCLUSION A wide spectrum of HPV genotypes is seen in this young population. Knowledge about HPV types prevalent in communities in different regions of India would be useful in devising the optimum strategy for cervical cancer prevention.

Bone marrow mononuclear cell therapy in ischaemic stroke: a systematic review

Bone marrow mononuclear cell (BM‐MNC) therapy has emerged as a potential therapy for the treatment of stroke. We performed a systematic review of published studies using BM‐MNC therapy in patients with ischaemic stroke (IS). Literature was searched using MEDLINE, PubMed, EMBASE, Trip Database, Cochrane library and clinicaltrial.gov to identify studies on BM‐MNC therapy in IS till June, 2016. Data were extracted independently by two reviewers. STATA version 13 was used for carrying out meta‐analysis. We included non‐randomized open‐label, single‐arm and non‐randomized comparative studies or randomized controlled trials (RCTs) if BM‐MNCs were used to treat patients with IS in any phase after the index stroke. One randomized trial, two non‐randomized comparative trials and four single‐arm open‐label trials (total seven studies) involving 227 subjects (137 patients and 90 controls) were included in the systematic review and meta‐analysis. The pooled proportion for favourable clinical outcome (modified Rankin Scale score ≤2) in six studies involving 122 subjects was 29% (95% CI 0.16–0.43) who were exposed to BM‐MNCs and pooled proportion for favourable clinical outcome of 69 subjects (taken from two trials) who did not receive BM‐MNCs was 20% (95% CI 0.12–0.32). The pooled difference in the safety outcomes was not significant between both the groups. Our systematic review suggests that BM‐MNC therapy is safe up to 1 year post‐intervention and is feasible; however, its efficacy in the case of IS patients is debatable. Well‐designed randomized controlled trials are required to provide more information on the efficacy of BM‐MNC transplantation in patients with IS.

Relationship of phosphodiesterase 4D (PDE4D) gene polymorphisms with risk of ischemic stroke: a hospital based case-control study

Abstract Background: Stroke remains a leading cause of death and disability worldwide. Ischemic stroke (IS) accounts for around 80–85% of total stroke and is a complex polygenic multi-factorial disorder which is affected by a complex combination of vascular, environmental, and genetic factors. Objective: The study was conducted with an aim to examine the relationship of single nucleotide polymorphisms (SNPs) of PDE4D (T83C, C87T, and C45T) gene with increasing risk of IS in patients in North Indian population. Methods: In this hospital-based case-control study, 250 IS subjects and 250 age-and sex-matched control subjects were enrolled from the Neurosciences Centre, A.I.I.M.S., New Delhi, India. Deoxyribonucleic acids (DNAs) were extracted using the conventional Phenol–Chloroform isolation method. Different genotypes were determined by Polymerase chain reaction– Restriction fragment length polymorphism method. Odds ratio (OR) and 95% Confidence Interval (CI) of relationship of polymorphisms with risk of IS were calculated by conditional multivariable regression analysis. Results: High blood pressure, low socioeconomic status, dyslipidemia, diabetes, and family history of stroke were observed to be statistically significant risk factors for IS. Multivariable adjusted analysis demonstrated a statistically significant relationship between SNP 83 of PDE4D gene polymorphism and increasing odds of IS under the dominant model of inheritance (OR, 1.59; 95% CI, 1.02 to 2.50; p value = 0.04) after adjustment of potential confounding variables. Stratified analysis on the basis of TOAST classification demonstrated a statistically significant association for increasing 2.73 times odds for developing large vessel disease stroke as compared to controls (OR, 2.73; 95% CI, 1.16 to 0.02; p value = 0.02). We did not find any significant association of SNPs (C87T and C45T) of the PDE4D gene with the risk of IS. Conclusion: SNP 83 of PDE4D gene may increase the risk for developing IS whereas SNP 87 and SNP45 of PDE4D may not be associated with the risk of IS in the North Indian population. Prospective cohort studies are required to corroborate these findings.

Abstract C59: Human papillomavirus genotype distribution in northern India: Implications for vaccination

Background: Cervical cancer caused by human papilloma virus (HPV), is the commonest cancer in Indian women, with an estimated 132000 new cases and 76000 deaths annually. More than 100 HPV genotypes have been identified in humans of which at least 40 HPV types are found in the genital tract. Several high‐risk HPV types are known to cause the disease. There are only limited data available on the genotype distributions in this region. Methods: During the period three different cross‐sectional studies were carried out: (1) a hospital‐based cross‐sectional study in which 106 women presenting with invasive cervical cancer; (2) a hospital‐based cross‐sectional study in which 524 women presenting to have an unhealthy cervix were invited to participate; (3) a community‐based cross‐sectional study in which 465 women also underwent HPV testing. HPV typing in these studies was done by using L1 consensus PCR followed by reverse line blot hybridization assay (Roche). Results: In the hospital‐based population, among 106 invasive cervical cancers the six commonest HPV types were HPV‐16, ‐18, ‐45, ‐59, ‐33 and 73. HPV‐16/18, singly or in combination, were seen in 83% of cases. (2) In the hospital‐based population, among the 524 symptomatic hospital subjects, the overall HPV prevalence was 15.5 % and the six commonest HPV types were as follows: HPV ‐16, ‐89, ‐39, ‐52, ‐33 and 18. Among the HPV positive women accounted for HPV‐16/18 were associated with 34.3%, 45.4% and 65.7% of normal, low‐grade and high‐grade disease respectively. Among (3) the community‐based women, the overall HPV prevalence was 7 % and the six commonest HPV types were as follows: HPV‐ 16, ‐ 45, ‐52, ‐31, ‐66, and ‐18. Conclusions: A wide spectrum of HPV types is seen in north Indian women, with HPV‐16/18 are the commonest types in all populations. However, thereafter there is considerable variation. It is estimated that the HPV ‐16/18 vaccine would reduce over 75% of the total cancer burden in this region. The community population is from a confined geographical area whereas the hospital population includes not only residents of Delhi and neighbouring areas but also further states of North India as well as neighboring countries of Nepal and Bangladesh. Thus more studies are required to establish the true picture of HPV type‐distribution in this large and diverse region. Citation Information: Cancer Res 2009;69(23 Suppl):C59.