Establishment of reverse transcription polymerase chain reaction assay for simultaneous detection of human parainfluenza virus types 1 to 4 in children with influenza like illnesses

M. GhelichKhan, S. M. Hosseini, B. Khansarinejad, F. Fotouhi
2018 Bratislava Medical Journal  
OBJECTIVES: The aim of this study was to develop an in-house multiplex reverse transcription polymerase chain reaction (mRT-PCR), which can recognize HPIV1-4 in clinical samples. BACKGROUND: Human parainfl uenza virus (HPIV) is one of the major causes of viral respiratory infections and can affect people at any age, especially infants and young children. METHODS: Four sets of specifi c primers targeting conserved areas of hemagglutinin-neuraminidase (HN) genes of HPIV1-4, were designed and
more » ... d with type-related plasmid controls. Specifi city and sensitivity of mPCR were tested. One-step mRT-PCR was set up using a viral panel containing 10 respiratory viruses, including HPIVs. One hundred nasopharyngeal samples of respiratory infection patients were tested using the set One-step mRT-PCR. RESULTS: The specifi city of set mPCR for HPIV1-4 using plasmid positive controls was proved and reaction sensitivity was measured. The specifi city of set mRT-PCR was confi rmed and 4 and 5 out of 100 clinical samples were HPIV1 and HPIV2 positive, respectively. CONCLUSION: The developed one-step mRT-PCR in this study is an effective and specifi c assay for clinical diagnosis of HPIV1 to 4 (Tab. 1, Fig. 6 , Ref. 28). Text in PDF www.elis.sk. KEY WORDS: human parainfl uenza virus, human, reverse transcriptase PCR, respiratory tract infections. 27. Fan J, Henrickson KJ. Rapid diagnosis of human parainfl uenza virus type 1 infection by quantitative reverse transcription-PCR-enzyme hybridization assay.
doi:10.4149/bll_2017_145 pmid:29322811 fatcat:744zb4wbvbg77igqjob3zmzngm