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, Wayne M. Sullender, Sushil K. Kabra, Kalaivani Mani, John Cherian, Vikas Tyagi, Harendra S. Chahar, Samander Kaushik, Lalit Dar, Shobha Broor

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

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 (PIV13) and human metapneumovirus (hMPV). Nasopharyngeal aspirates (NPAs) were collected from children with ALRI 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 2005March 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.

Original languageEnglish (US)
Article number89
JournalVirology Journal
Volume6
DOIs
StatePublished - 2009
Externally publishedYes

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Multiplex Polymerase Chain Reaction
Urban Hospitals
Virus Diseases
Respiratory Tract Infections
Metapneumovirus
Pediatrics
Viruses
Respiratory Syncytial Viruses
Indirect Fluorescent Antibody Technique
Centrifugation
Human Influenza
India
Paramyxoviridae Infections
Respiratory Syncytial Virus Infections
Polymerase Chain Reaction
Bronchiolitis
Orthomyxoviridae
Coinfection
Outpatients
Morbidity

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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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. / Bharaj, Preeti; Sullender, Wayne M.; Kabra, Sushil K.; Mani, Kalaivani; Cherian, John; Tyagi, Vikas; Chahar, Harendra S.; Kaushik, Samander; Dar, Lalit; Broor, Shobha.

In: Virology Journal, Vol. 6, 89, 2009.

Research output: Contribution to journalArticle

Bharaj, Preeti ; Sullender, Wayne M. ; Kabra, Sushil K. ; Mani, Kalaivani ; Cherian, John ; Tyagi, Vikas ; Chahar, Harendra S. ; Kaushik, Samander ; Dar, Lalit ; Broor, Shobha. / 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. In: Virology Journal. 2009 ; Vol. 6.
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abstract = "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 (PIV13) and human metapneumovirus (hMPV). Nasopharyngeal aspirates (NPAs) were collected from children with ALRI 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 2005March 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.",
author = "Preeti Bharaj and Sullender, {Wayne M.} and Kabra, {Sushil K.} and Kalaivani Mani and John Cherian and Vikas Tyagi and Chahar, {Harendra S.} and Samander Kaushik and Lalit Dar and Shobha Broor",
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T1 - 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

AU - Bharaj, Preeti

AU - Sullender, Wayne M.

AU - Kabra, Sushil K.

AU - Mani, Kalaivani

AU - Cherian, John

AU - Tyagi, Vikas

AU - Chahar, Harendra S.

AU - Kaushik, Samander

AU - Dar, Lalit

AU - Broor, Shobha

PY - 2009

Y1 - 2009

N2 - 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 (PIV13) and human metapneumovirus (hMPV). Nasopharyngeal aspirates (NPAs) were collected from children with ALRI 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 2005March 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.

AB - 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 (PIV13) and human metapneumovirus (hMPV). Nasopharyngeal aspirates (NPAs) were collected from children with ALRI 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 2005March 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.

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