Combined SYBR Green real-time polymerase chain reaction and microarray method for the simultaneous determination of human papillomavirus loads and genotypes

Hyun Hee Seo; Young Jun Kim; Mi Seon Jeong; 홍성란; In-Ho Lee; So Kyeong A; 김미경; 이유경; Ki-Heon Lee; 김주리; 김성재; 김태진

Combined SYBR Green real-time polymerase chain reaction and microarray method for the simultaneous determination of human papillomavirus loads and genotypes

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Type: Academic journal

Author: Hyun Hee Seo Young Jun Kim Mi Seon Jeong 홍성란 In-Ho Lee So Kyeong A 김미경 이유경 Ki-Heon Lee 김주리 김성재 김태진

Journal: Korean Society of Obstetrics and Gynecology Obstetrics & Gynecology Science Obstetrics & Gynecology Science 제59권 제6호 KCI Accredited Journals

Published: 2016.1

Pages: 489 - 497 (9page)

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Combined SYBR Green real-time polymerase chain reaction and microarray method for the simultaneous determination of human papillomavirus loads and genotypes

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ObjectiveThe aim of this study was to describe the principle of the Cheil HPV DNA Chip assay and evaluate its accuracy. In order to quantify the human papillomavirus (HPV) load and identify HPV genotypes simultaneously, this assay combined the two methods: SYBR Green quantitative real-time polymerase chain reaction (PCR) and DNA microarray. MethodsWe designed novel consensus primer sets that target the conserved region of the HPV L1 gene for quantifying and detecting a broad range of HPV types by quantitative real-time PCR. Subsequently, using the PCR products, DNA microarray was performed with 36 HPV type-specific probes. To validate this method, direct sequencing and correlation analysis among HPV genotype, viral load, and cytological abnormality was performed by Cohen’s kappa values, two-sided McNemar chi-square test, Kruskal-Wallis test, and odds ratios. ResultsThe kappa value of the Cheil HPV DNA Chip was 0.963 (95% confidence interval, 0.919 to 0.98), which was significantly higher than the value of 0.527 (95% confidence interval, 0.447 to 0.59) obtained using a conventional HPV DNA Chip. HPV16 (χ2=62.28, P<0.01), HPV33 (χ2=7.18, P<0.01), and HPV58 (χ2=9.52, P<0.01), which are classified as high-risk HPVs, were detected at significant levels in samples with high-grade lesions. And viral loads tended to be higher in groups with high odds ratios. ConclusionThe Cheil HPV DNA Chip is an effective diagnostic assay for simultaneously detecting HPV genotypes and loads in cervical samples.

#Diagnosis #Genotyping #Human papilloma virus #Microarray #Viral load

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Dalstein V / 2003 / Persistence and load of highrisk HPV are predictors for development of high-grade cervical lesions : a longitudinal French cohort study / Int J Cancer 106:396~403

Schlecht NF / 2001 / Persistent human papillomavirus infection as a predictor of cervical intraepithelial neoplasia / JAMA 286:3106~3114

Lorincz AT / 1992 / Human papillomavirus infection of the cervix : relative risk associations of 15 common anogenital types / Obstet Gynecol 79:328~337

Walboomers JM / 1999 / Human papillomavirus is a necessary cause of invasive cervical cancer worldwide / J Pathol 189:12~19

Heard I / 2000 / Increased risk of cervical disease among human immunodeficiency virus-infected women with severe immunosuppression and high human papillomavirus load / Obstet Gynecol 96:403~409

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