A duplex RT-PCR for identifying genotypes of human metapneumovirus from clinical samples
10.3760/cma.j.issn.1009-9158.2010.01.008
- VernacularTitle:人偏肺病毒基因型双重逆转录PCR检测方法的建立
- Author:
Fang WANG
;
Runan ZHU
;
Yuan QIAN
;
Jie DENG
;
Yu SUN
;
Linqing ZHAO
;
Bin LIAO
;
Rongyan HUANG
- Publication Type:Journal Article
- Keywords:
Metapneumovirus;
Genotype;
Reverse transcriptase polymerase chain reaction
- From:
Chinese Journal of Laboratory Medicine
2010;33(1):33-36
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a convenient reverse transcription PCR(RT-PCR)method for identifying genotypes of human metapneumovirus(hMPV)from clinical samples.Methods According to the gene sequences of hMPV G with different genotypes,the A and B genotype specific primers were designed.A diplex RT-PCR was applied to identify different genotypes according to the molecular weight of PCR products in agarose gel.37 clinical samples were detected through this method.Results It was convenient to distinguish different genotypes of hMPV(383 bp for A and 284 bp for B)by the diplex RTPCR,and there was no non-specific amplification for common respiratory viruses.so it meant that the specificity of primers was good.The results of genotyping 37 clinical samples showed that 20 samples were identified as genotype A by both sequence analysis of M gene and diplex RT-PCR,whereas 17 samples were identified as genotype B by sequence analysis of M gene.but in these 17 samples 14 samples were identified as genotype B by the diplex RT-PCR and remaining 3 samples could not be genotyped because there was no PCR product after amplification.The consistency rate for these two methods Was 91.9%[(20+14)/37].Conclusion The method of diplex RT-PCR Was developed successfully and can be used for identify genotypes of hMPV.
