Isolation & molecular characterization of human parainfluenza virus in Chennai, India

Abstract

Background & objectives: Human parainfluenza virus (HPIV) accounts for a significant proportion of lower respiratory tract infections in children as well as adults. This study was done to detect the presence of different subtypes of HPIV from patients having influenza like illness (ILI).

Methods: Throat and nasal swabs from 232 patients with ILI who were negative for influenza viruses were tested by multiplex reverse transcription polymerase chain reaction(mRT-PCR) for the detection of human parainfluenza virus. All samples were inoculated in rhesus monkey kidney (LLC-MK2) cell line.

Results: Of the 232 samples, 26(11.2%) were positive by mRT-PCR and nine (34.6%) showed cytopathic effect with syncytium formation for HPIV and all were HPIV-3 serotype, other serotypes like 1,2,4 were negative. The HPIV-3 strains (HN gene) were sequenced and analysed. Two novel mutations were identified at amino acid residues 295 and 297.

Interpretation & conclusions: The mRT-PCR assay offers a rapid, sensitive and accurate diagnostic method for detection of HPIV which enables early detection and control. In our study there was a predominance of HPIV among 1-5 yr age group and the school going age group was less affected. Further studies need to be done to characterize HPIV isolated from different parts of the country.

Fig. 1

Results of multiplex reverse-transcription (RT)-PCR with clinical samples and controls. Lane M, Marker; Lane 1, HPIV-1(Positive control); Lane 2, HPIV-2(PC); Lane 3, HPIV-3(PC); Lane 4, HPIV4(PC); Lane 5, Negative; Lane 6, HPIV3(clinical sample); Lanes 7 to 11, Negative; Lane 12, HPIV-3 (clinical sample); Lane 13, Negative control

Fig. 2

Phylogenetic analysis of the deduced amino acid sequences of hemagglutinin neuraminidase gene of HPIV-3 to members of the family Paramyxoviridae. The tree was constructed by the maximum likelihood method with p distance.