基于AllGlo探针的双重实时荧光定量PCR法检测高危型人乳头瘤病毒HPV16/18

Simultaneous detection of high-risk Human papillomaviruses 16/18 by AllGlo probe-based multiplex quantitative real-time PCR

目的建立基于AllGlo探针的双重实时荧光定量PCR检测高危型人乳头瘤病毒HPV16/18的方法。方法针对HPV16/18病毒基因组保守序列,设计引物和AllGlo探针,建立双重荧光定量PCR检测方法,评价其敏感性、特异性和稳定性,并与传统HC2方法比较。结果成功建立了双重实时荧光定量PCR检测HPV16/18的方法,检测限均为10copies/μL,敏感性和特异性均为100%。应用该方法检测160份宫颈肿瘤患者样本,与传统HC2法结果一致性为100%。结论该研究建立的快速、准确、可同时检测HPV16/18的双重实时荧光定量PCR法,具有较高的敏感性、特异性和稳定性,操作简单,可用于HPV16/18的临床筛查和诊断。

Objective To develop an AIlGlo probe-based multiplex real-time PCR assay for simultaneous detection of 2 high- risk Human papillomaviruses （HPV） HPV16/HPV18. Methods According to conserved regions of HPV16/18, specific primers and probes were designed. The assay was established after optimization, sensitivity, specificity and the stability of the assay was evaluated. The assay was applied for analysis of clinical specimens; results were compared to those of traditional as- say HC2. Results AllGlo probe-based multiplex PCR for detection of HPV16/18 was successfully established, limit of detec- tion was 10 copies/μl for both HPV16 and HPV18. Sensitivity and specificity was both 100%. 160 clinical samples collected from patients with cervical cancer were tested by the assay, result was 100% consistent with traditional HC2 method. Conclu- sion AllGlo probe-based multiplex real-time PCR established in this study could simultaneously detect HPV16/18 in a quick and accurate manner, with high sensitivity, specificity and stability. In addition, it is easy to operate to make it applicable for clinical screening and diagnosis.