Detection and subgrouping of respiratory syncytial virus directly from nasopharyngeal aspirates

Reena Ghildyal, Geoff Hogg, John Mills, Jayesh Meanger

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

OBJECTIVE: To develop a reverse transcription-polymerase chain reaction (RT-PCR)-based assay to identify the subgroup of the infecting respiratory syncytial virus (RSV) strain directly from nasopharyngeal aspirates (NPAs). METHODS: A total of 154 NPAs which were positive for RSV antigen by direct immunofluorescence were subjected to RT-PCR. The primers used were designed to give products for subgroup A and B which were of different sizes and easily visualized on agarose electrophoresis. The PCR products were further analyzed by restriction analysis using enzymes which were unique or rare cutters within the PCR amplimer. RESULTS: It was possible to confirm RSV infection in 70% of the NPA samples studied. Of these, 92.6% belonged to the A group, and only 7.4% to the B group. Within the A group, six subgroups were identified using restriction analysis, while all B-group samples were identical to the prototype B strain, 18537. CONCLUSION: RT-PCR performed on RNA isolated directly from NPAs provides a quick, easy-to-use, reasonably sensitive method to identify and group the infecting RSV strain.

Original languageEnglish
Pages (from-to)120-123
Number of pages4
JournalClinical Microbiology and Infection
Volume3
Issue number1
DOIs
Publication statusPublished - Feb 1997
Externally publishedYes

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Respiratory Syncytial Viruses
Polymerase Chain Reaction
Reverse Transcription
Respiratory Syncytial Virus Infections
Direct Fluorescent Antibody Technique
Restriction Mapping
Sepharose
Electrophoresis
RNA
Antigens

Cite this

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title = "Detection and subgrouping of respiratory syncytial virus directly from nasopharyngeal aspirates",
abstract = "OBJECTIVE: To develop a reverse transcription-polymerase chain reaction (RT-PCR)-based assay to identify the subgroup of the infecting respiratory syncytial virus (RSV) strain directly from nasopharyngeal aspirates (NPAs). METHODS: A total of 154 NPAs which were positive for RSV antigen by direct immunofluorescence were subjected to RT-PCR. The primers used were designed to give products for subgroup A and B which were of different sizes and easily visualized on agarose electrophoresis. The PCR products were further analyzed by restriction analysis using enzymes which were unique or rare cutters within the PCR amplimer. RESULTS: It was possible to confirm RSV infection in 70{\%} of the NPA samples studied. Of these, 92.6{\%} belonged to the A group, and only 7.4{\%} to the B group. Within the A group, six subgroups were identified using restriction analysis, while all B-group samples were identical to the prototype B strain, 18537. CONCLUSION: RT-PCR performed on RNA isolated directly from NPAs provides a quick, easy-to-use, reasonably sensitive method to identify and group the infecting RSV strain.",
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Detection and subgrouping of respiratory syncytial virus directly from nasopharyngeal aspirates. / Ghildyal, Reena; Hogg, Geoff; Mills, John; Meanger, Jayesh.

In: Clinical Microbiology and Infection, Vol. 3, No. 1, 02.1997, p. 120-123.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Detection and subgrouping of respiratory syncytial virus directly from nasopharyngeal aspirates

AU - Ghildyal, Reena

AU - Hogg, Geoff

AU - Mills, John

AU - Meanger, Jayesh

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N2 - OBJECTIVE: To develop a reverse transcription-polymerase chain reaction (RT-PCR)-based assay to identify the subgroup of the infecting respiratory syncytial virus (RSV) strain directly from nasopharyngeal aspirates (NPAs). METHODS: A total of 154 NPAs which were positive for RSV antigen by direct immunofluorescence were subjected to RT-PCR. The primers used were designed to give products for subgroup A and B which were of different sizes and easily visualized on agarose electrophoresis. The PCR products were further analyzed by restriction analysis using enzymes which were unique or rare cutters within the PCR amplimer. RESULTS: It was possible to confirm RSV infection in 70% of the NPA samples studied. Of these, 92.6% belonged to the A group, and only 7.4% to the B group. Within the A group, six subgroups were identified using restriction analysis, while all B-group samples were identical to the prototype B strain, 18537. CONCLUSION: RT-PCR performed on RNA isolated directly from NPAs provides a quick, easy-to-use, reasonably sensitive method to identify and group the infecting RSV strain.

AB - OBJECTIVE: To develop a reverse transcription-polymerase chain reaction (RT-PCR)-based assay to identify the subgroup of the infecting respiratory syncytial virus (RSV) strain directly from nasopharyngeal aspirates (NPAs). METHODS: A total of 154 NPAs which were positive for RSV antigen by direct immunofluorescence were subjected to RT-PCR. The primers used were designed to give products for subgroup A and B which were of different sizes and easily visualized on agarose electrophoresis. The PCR products were further analyzed by restriction analysis using enzymes which were unique or rare cutters within the PCR amplimer. RESULTS: It was possible to confirm RSV infection in 70% of the NPA samples studied. Of these, 92.6% belonged to the A group, and only 7.4% to the B group. Within the A group, six subgroups were identified using restriction analysis, while all B-group samples were identical to the prototype B strain, 18537. CONCLUSION: RT-PCR performed on RNA isolated directly from NPAs provides a quick, easy-to-use, reasonably sensitive method to identify and group the infecting RSV strain.

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