Llorente J L, Hidalgo F I, Melón S, de Oña M, Carreño M, Suárez C
ENT Department, Hospital Central de Asturias Universidad de Oviedo, Spain.
Auris Nasus Larynx. 1998 Dec;25(4):387-92. doi: 10.1016/s0385-8146(98)00038-8.
Nested polymerase chain reaction (nested PCR) was performed using a reaction mix batch-prepared and kept frozen in single reaction tubes at -20 degrees C until use. Twenty-one New Zealand white rabbits were infected with herpes simplex virus type 1 (HSV-1). Eleven animals were killed on day seven and the other ten were sacrificed on day 21. Viral culture and nested PCR was used to determine the presence of HSV-1 in samples from the tongue, HSV-1 was detected in 90.47% of the animals; in 84.21% by nested PCR and in 52.63% by culture. Nested PCR assay had greater sensitivity than culture in animals sacrificed on day seven with significative difference (p < 0.05). Higher sensitivity and faster results were obtained with this method, so we found it reliable and useful in the setting of a clinical laboratory dealing with diagnosis of herpes virus infections.
采用巢式聚合酶链反应（nested PCR），反应混合物预先批量制备并保存在单个反应管中，于-20℃冷冻，直至使用。21只新西兰白兔感染了1型单纯疱疹病毒（HSV-1）。11只动物在第7天处死，另外10只在第21天处死。采用病毒培养和巢式PCR检测舌部样本中HSV-1的存在情况，90.47%的动物检测到HSV-1；巢式PCR检测阳性率为84.21%，病毒培养检测阳性率为52.63%。在第7天处死的动物中，巢式PCR检测的灵敏度高于病毒培养，差异有统计学意义（p < 0.05）。该方法具有更高的灵敏度和更快的检测结果，因此我们发现它在临床实验室诊断疱疹病毒感染时可靠且有用。
