Zhong Q, Li K, Chen D, Wang H, Lin Q, Liu W
a Department of Central Laboratory , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China.
b Department of Laboratory Medicine , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China.
Br J Biomed Sci. 2018 Jul;75(3):110-115. doi: 10.1080/09674845.2017.1411864. Epub 2018 Mar 14.
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.
目的 尖锐湿疣（CA）是一种常见的、由病毒引起的全球性性传播疾病。人乳头瘤病毒（HPV）基因分型对CA的治疗具有重要临床意义。我们开发了一种环介导等温扩增（LAMP）方法用于检测HPV。方法 我们收集了294份宫颈刮片样本，包括30份HPV-6阳性、30份HPV-11阳性、22份HPV-16阳性、20份HPV-42阳性、30份HPV-43阳性、20份HPV-44阳性以及142份HPV阴性样本。对40例经病理诊断为CA的患者的组织进行石蜡包埋，并采用LAMP和Luminex技术进行分析。使用羟基萘酚蓝（HNB）和电泳检测LAMP结果。结果 对LAMP和Luminex系统检测HPV六种亚型的情况进行了比较。LAMP反应对每种亚型具有特异性。通过HNB指示剂法测定，LAMP对HPV-6的灵敏度为1000拷贝/管。两种方法之间的kappa值分别为：HPV-6为0.98，HPV-11为0.94，HPV-43为0.89，HPV-42为0.87，HPV-16为0.79，HPV-44为0.68。在Luminex检测确定的142份HPV阴性样本中，LAMP检测出8份HPV-6阳性和1份HPV-11阳性。在40份CA样本中，LAMP和Luminex的结果在38份（95%）样本中一致。结论 本研究结果表明，采用HNB的LAMP检测法在灵敏度和特异性方面优于Luminex方法。LAMP的特异性为100%，使用HNB时LAMP的灵敏度为1000拷贝/管。因此，LAMP是一种用于临床诊断HPV亚型的有用、快速且准确的方法。
