实时荧光PCR方法快速检测KI多瘤病毒

Rapid identification of KI polyomaviruses in clinical samples by real-time PCR

目的采用实时荧光PCR方法快速检测KI多瘤病毒(KIPy V),并与巢式PCR扩增法进行比较。方法收集2007年11月至2015年1月在福州市一家妇幼保健院因呼吸道感染住在儿科重症监护病房(PICU)的259例小儿鼻咽抽取物标本和另两家综合医院146例因严重急性呼吸道感染(SARI)住院的成年患者咽拭子标本,通过实时荧光PCR方法对KIPy V的调节区的基因片段进行检测。结果在小儿鼻咽抽取物标本中检测出3例KIPy V感染阳性病例,检测阳性率约为1.2%,而用巢式PCR扩增法只检测出1例,这例的Ct值较低,约为23,并且扩增曲线呈S型。其它两例的Ct值较高,分别约为33和35。在成年患者咽拭子标本中,用两种方法都未检测出KIPy V感染。在两例Ct值较高的小儿病例中,检测出混合有呼吸道合胞病毒(RSV)感染。结论确立的实时荧光PCR方法可以快速检测KIPy V,且特异性较高。

Objective To develop sensitive and specific assays for the detection of KI polyomavirus（KIPy V）. Methods In our study,real-time polymerase chain reaction（rt PCR） assays were developed and compared with nested PCR for detecting KIPy V. Nasopharyngeal aspirates（NPA） were collected from children with respiratory tract disease from Nov. 2007 to Jan. 2015. Throat swabs were collected from SARI（serious acute respiratory infection） patients from 2013 to 2015. A total of 259 NPA samples and 146 throat swabs were tested for KIPy V using two PCR methods. Results KIPy V infection was detected in 3 NPA samples by rt PCR assay. The positive rate was 1.2%. The Ct value of which was detected by nested PCR was about 23 by rt PCR assay. The Ct values of the other 2 samples were about 33 and 35,and both of them were not detected by nested PCR. Conclusion RT-PCR is a sensitive and fast detection assay for KIPy V in clinical specimens and useful for further research of this virus.