[Development of a SYBR green real-time RT-PCR for detection of mumps virus gene].
OBJECTIVE
A SYBR Green real-time RT-PCR assay was developed to detect mumps virus gene rapidly.


METHODS
The primers were selected based on SH gene of mumps virus, the assay was optimized in reactive system and condition to improve the sensitivity and specificity. The SH gene of 5 mumps virus isolates were detected and genotyped using this assay.


RESULTS
A SYBR Green real-time RT-PCR assay had good sensitivity and specificity. There was no cross reaction with other respiratory virus. The detection limit of the assay was 10 TCID50. The sensitivity of the two assays, RT-PCR and real-time RT-PCR, was not different. Compared with traditional RT-PCR, real-time RT-PCR saved half detection time. The products of SYBR Green real-time RT-PCR were directly sequenced and genotyped.


CONCLUSIONS
SYBR Green real-time RT-PCR was a rapid, specific and sensitive method for the detection of mumps virus and genotyping.
Cui
Ai-Li
AL
State Key Laboratory for Molecular Virology and Genetic, Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Zhu
Zhen
Z
Wang
Chang-Yin
CY
D052160:Benzothiazoles; D003959:Diamines; D006801:Humans; D008969:Molecular Sequence Data; D009107:Mumps / Q000821:virology*; D009109:Mumps virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D009930:Organic Chemicals / Q000378:metabolism; D010802:Phylogeny; D011804:Quinolines; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000235:genetics*
D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D014764:Viral Proteins; C098022:SYBR Green I