title: A novel duplex real time quantitative reverse transcription polymerase chain reaction for rubella virus with armored RNA as a noncompetitive internal positive control.
abstract: The objective of this study was to build and apply a duplex real time quantitative reverse transcription-polymerase chain reaction (RT-PCR) for rubella virus. Firstly, a 60-bp-long armored RV RNA was constructed in the laboratory. Secondly, a duplex real time RT-PCR assay was established. Thirdly, the 60-bp-long armored RV RNA was used as an internal positive control (IPC) for the duplex real time RT-PCR. And finally the duplex real time RT-PCR assay was applied to detect RV RNA in clinical specimens. The in-house assay has a high amplification efficiency (0.99), a high analytical sensitivity (200 copies/mL), and a good reproducibility. The diagnostic specificity and sensitivity of the in-house assay were both 100%, due to the monitoring of the armored RV RNA IPC. Therefore, the in-house duplex real time quantitative RT-PCR assay is a specific, sensitive, reproducible and accurate assay for quantitation of RV RNA in clinical specimens. And noncompetitive armored RV RNA IPC can monitor RT-PCR inhibition and prevent false-negative and inaccurate results in the real time detection system.
authors: Department of Laboratory, Tai'an Central Hospital, Tai'an 271000, China.
authors: Lihong
authors: L
authors: Zhao
authors: Department of Reproductive Genetics, Tai'an Central Hospital, Tai'an 271000, China.
authors: Ruiying
authors: R
authors: Li
authors: Central Laboratory, Tai'an Central Hospital, Tai'an 271000, China.
authors: Aihua
authors: A
authors: Liu
authors: Department of Laboratory, Tai'an Central Hospital, Tai'an 271000, China. Electronic address: zhaolihong70@163.com.
authors: Shuping
authors: S
authors: Zhao
mesh_terms: D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D015203:Reproducibility of Results; D012409:Rubella / Q000175:diagnosis* / Q000821:virology*; D012412:Rubella virus / Q000235:genetics*; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D012367:RNA, Viral
keywords: Armored RNA; Internal positive control; Real time RT-PCR; Rubella virus