Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2009 Jun 26:6:89.
doi: 10.1186/1743-422X-6-89.

Respiratory viral infections detected by multiplex PCR among pediatric patients with lower respiratory tract infections seen at an urban hospital in Delhi from 2005 to 2007

Preeti Bharaj  1 , Wayne M Sullender, Sushil K Kabra, Kalaivani Mani, John Cherian, Vikas Tyagi, Harendra S Chahar, Samander Kaushik, Lalit Dar, Shobha Broor
Affiliations
Comparative Study

Respiratory viral infections detected by multiplex PCR among pediatric patients with lower respiratory tract infections seen at an urban hospital in Delhi from 2005 to 2007

Preeti Bharaj et al. Virol J. .

Abstract

Background: Acute lower respiratory tract infections (ALRI) are the major cause of morbidity and mortality in young children worldwide. Information on viral etiology in ALRI from India is limited. The aim of the present study was to develop a simple, sensitive, specific and cost effective multiplex PCR (mPCR) assay without post PCR hybridization or nested PCR steps for the detection of respiratory syncytial virus (RSV), influenza viruses, parainfluenza viruses (PIV1-3) and human metapneumovirus (hMPV). Nasopharyngeal aspirates (NPAs) were collected from children with ALRI < or = 5 years of age. The sensitivity and specificity of mPCR was compared to virus isolation by centrifugation enhanced culture (CEC) followed by indirect immunofluorescence (IIF).

Results: From April 2005-March 2007, 301 NPAs were collected from children attending the outpatient department or admitted to the ward of All India Institute of Medical Sciences hospital at New Delhi, India. Multiplex PCR detected respiratory viruses in 106 (35.2%) of 301 samples with 130 viruses of which RSV was detected in 61, PIV3 in 22, PIV2 in 17, hMPV in 11, PIV1 in 10 and influenza A in 9 children. CEC-IIF detected 79 viruses only. The sensitivity of mPCR was 0.1TCID50 for RSV and influenza A and 1TCID50 for hMPV, PIV1, PIV2, PIV3 and Influenza B. Mixed infections were detected in 18.8% of the children with viral infections, none detected by CEC-IIF. Bronchiolitis was significantly associated with both total viral infections and RSV infection (p < 0.05). History of ARI in family predisposed children to acquire viral infection (p > 0.05).

Conclusion: Multiplex PCR offers a rapid, sensitive and reasonably priced diagnostic method for common respiratory viruses.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Standardization of two tube multiplex PCR for RSV, Influenza A&B viruses in first tube and PIV1–3 and hMPV in the second tube. Lane 1: Marker Ø X174 (Hae III digested). Lane 2: Amplicon forRSV showing band of 683 bp, Influenza A of 105 bp, Influenza B of 503 bp. Lane 3: Marker 100 bp ladder. Lane 4: Amplicon for PIV1 showing band of 84 bp, PIV2 of 197 bp, PIV3 of 266 bp, and hMPV of 440 bp.
Figure 2
Figure 2
Application of two tube multiplex PCR on clinical samples. Panel A. Lane 1: 100 bp DNA ladder. Lane 2: Clinical sample showing amplicon of 105 bp for FLU A. Lane 3: Clinical sample showing amplicon of 683 bp for RSV. Lane 4: Clinical sample showing amplicon of 440 bp for hMPV. Lane 5: Clinical sample showing amplicon of 266 bp for PIV3. Lane 3: Negative clinical sample. Panel B. Lane 1: 100 bp DNA adder. Lane 2: Clinical sample showing mixed infection of PIV1 (84 bp), PIV2 (197 bp) and PIV3 (266 bp).
Figure 3
Figure 3
Monthly distribution of ALRI causing viruses detected during the study.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Hijazi Z, Pacsa A, Eisa S, el Shazli A, abd el-Salam RA. Laboratory diagnosis of acute lower respiratory tract viral infections in children. J Trop Pediatr. 1996;42:276–80. doi: 10.1093/tropej/42.5.276. - DOI - PubMed
    1. Hinman AR. Global progress in infectious disease control. Vaccine. 1998;16:1116–21. doi: 10.1016/S0264-410X(98)80107-2. - DOI - PubMed
    1. Glezen WP, Loda FA, Clyde WA, Jr, Senior RJ, Sheaffer CI, Conley WG, Denny FW. Epidemiologic patterns of acute lower respiratory disease of children in a pediatric group practice. J Pediatr. 1971;78:397–406. doi: 10.1016/S0022-3476(71)80218-4. - DOI - PubMed
    1. Fan J, Henrickson KJ, Savatski LL. Rapid simultaneous diagnosis of infections with respiratory syncytial viruses A and B, influenza viruses A and B, and human parainfluenza virus types 1, 2, and 3 by multiplex quantitative reverse transcription-polymerase chain reaction-enzyme hybridization assay (Hexaplex) Clin Infect Dis. 1998;26:1397–1402. doi: 10.1086/516357. - DOI - PubMed
    1. Bellau-Pujol S, Vabret A, Legrand L, Dina J, Gouarin S, Petitjean-Lecherbonnier J, Pozetto B, Ginevra C, Freymuth F. Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses. J Virol Methods. 2005;126:53–63. doi: 10.1016/j.jviromet.2005.01.020. - DOI - PMC - PubMed

Publication types