Rapid detection and subtyping of human papillomaviruses in condyloma acuminatum using loop-mediated isothermal amplification with hydroxynaphthol blue dye
Rapid detection and subtyping of human papillomaviruses in condyloma acuminatum using loop-mediated isothermal amplification with hydroxynaphthol blue dye
复制标题
使用羟基萘酚蓝染料环介导等温扩增快速检测尖锐湿疣中的人乳头瘤病毒并进行分型
DOI：
10.1080/09674845.2017.1411864
复制
发表时间：
2018-01-01
影响因子：
1.9
通讯作者：
Liu, W.
中科院分区：
文献类型：
--
作者：
Zhong, Q.;Li, K.;Liu, W.
摘要目的尖锐湿疣（Condyloma acuminatum，CA）是一种常见的病毒性病.人乳头瘤病毒（HPV）基因分型对CA的治疗具有重要的临床意义。我们建立了一种检测HPV的环介导等温扩增（LAMP）方法。方法收集294例宫颈刮片标本，其中HPV-6阳性30例，HPV-11阳性30例，HPV-16阳性22例，HPV-42阳性20例，HPV-43阳性30例，HPV-44阳性20例，HPV阴性142例。对40例病理诊断为CA的患者的组织进行石蜡包埋，并通过LAMP和Luminex进行分析。采用羟基萘酚蓝（HNB）染色和电泳检测LAMP结果。结果LAMP和Luminex系统对6种HPV亚型的检测结果进行了比较。LAMP反应对每个亚型都有特异性。通过HNB指示剂测定，LAMP对HPV-6的灵敏度为1000拷贝/管。两种方法的Kappa值分别为0.98（HPV-6）、0.94（HPV-11）、0.89（HPV-43）、0.87（HPV-42）、0.79（HPV-16）和0.68（HPV-44）。在Luminex检测的142份HPV阴性样本中，LAMP检测到8份HPV-6和1份HPV-11。在40例CA标本中，LAMP法与Luminex法检测结果一致的有38例（95%）。结论本研究结果表明，在敏感性和特异性方面，HNB LAMP法优于Luminex法，具有上级优势。LAMP的特异性为100%，敏感性为1000拷贝/管。因此，LAMP是一种有用的，快速和准确的方法，用于HPV亚型的临床诊断。
Abstract Objective Condyloma acuminatum (CA) is a common, viral, sexually transmitted disease worldwide. Human papillomavirus (HPV) genotyping has important clinical implications for the treatment of CA. We developed a loop-mediated isothermal amplification (LAMP) method for the detection of HPV. Methods We collected 294 cervical scrape samples, including 30 HPV-6-positive, 30 HPV-11-positive, 22 HPV-16-positive, 20 HPV-42-positve, 30 HPV-43-positive, 20 HPV-44-positive and 142 HPV-negative samples. Tissues from 40 patients with a pathological diagnosis of CA were paraffin-embedded and analyzed by LAMP and Luminex. Hydroxynaphthol blue (HNB) and electrophoresis were used to detect the results of LAMP. Results LAMP and Luminex systems were compared in detecting six subtypes of HPV. LAMP reactions were specific for each subtype. The sensitivity of LAMP for HPV-6, as determined by the HNB indicator assay, was 1000 copies/tube. The kappa value between the two methods was 0.98 (HPV-6), 0.94 (HPV-11), 0.89 (HPV-43), 0.87 (HPV-42) 0.79 (HPV-16) and 0.68 (HPV-44). Among the 142 HPV-negative samples determined by the Luminex assay, HPV-6 was detected in eight and HPV-11 in one by LAMP. Among the 40 CA samples, the results of LAMP and Luminex were in agreement in 38 (95%). Conclusion The results of this study indicated that the LAMP assay with HNB is superior to the Luminex method in terms of sensitivity and specificity. The specificity of LAMP was 100% and the sensitivity of LAMP was 1000 copies/tube using HNB. LAMP is therefore a useful, quick and accurate method for the clinical diagnosis of HPV subtypes.