Detection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCR

Aguilar, Jose C.Perez-Breña, PilarGarcía, M LCruz, NErdman, D DEchevarria, Juan Emilio2019-05-312019-05-312000-03J Clin Microbiol. 2000 Mar;38(3):1191-5.0095-1137http://hdl.handle.net/20.500.12105/7704Correction. Detection and Identification of Human Parainfluenza Viruses 1, 2, 3, and 4 in Clinical Samples of Pediatric Patients by Multiplex Reverse Transcription-PCR. J Clin Microbiol 2000 Jul;38(7):2805. https://doi.org/10.1128/JCM.38.7.2805-2805.2000We describe a multiplex reverse transcription-PCR (m-RT-PCR) assay that is able to detect and differentiate all known human parainfluenza viruses (HPIVs). Serial dilution experiments with reference strains that compared cell culture isolation and m-RT-PCR showed sensitivities ranging from 0.0004 50% tissue culture infective dose (TCID(50)) for HPIV type 4B (HPIV-4B) to 32 TCID(50)s for HPIV-3. As few as 10 plasmids containing HPIV PCR products could be detected in all cases. When 201 nasopharyngeal aspirate specimens from pediatric patients hospitalized for lower respiratory illness were tested, m-RT-PCR assay detected 64 HPIVs (24 HPIV-3, 23 HPIV-1, 10 HPIV-4, and 7 HPIV-2), while only 42 of them (21 HPIV-1, 14 HPIV-3, 6 HPIV-2, and 1 HPIV-4 isolates) grew in cell culture. Our m-RT-PCR assay was more sensitive than either cell culture isolation or indirect immunofluorescence with monoclonal antibodies for the detection of HPIV infections. Also, HPIV-4 was more frequently detected than HPIV-2 in this study, suggesting that it may have been underestimated as a lower respiratory tract pathogen because of the insensitivity of cell culture.engAMhttp://creativecommons.org/licenses/by-nc-sa/4.0/AnimalsCell LineChildDNA PrimersDNA, ViralDogsFluorescent Antibody Technique, IndirectHumansInhalationParainfluenza Virus 1, HumanParainfluenza Virus 2, HumanParainfluenza Virus 3, HumanRespirovirus InfectionsRetrospective StudiesReverse Transcriptase Polymerase Chain ReactionRubulavirusRubulavirus InfectionsDetection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCRAtribución-NoComercial-CompartirIgual 4.0 Internacional106990203831191-5Journal of clinical microbiologyopen access