Comparison of Quantitative Reverse Transcription-PCR to Viral Culture for Assessment of Respiratory Syncytial Virus Shedding
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| Published in | Journal of Clinical Microbiology Vol. 41; no. 9; pp. 4160 - 4165 |
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American Society for Microbiology
01.09.2003
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Respiratory syncytial virus (RSV) has recently been recognized as a serious pathogen in elderly and immunocompromised adults. Diagnosis of acute infection in adults is often difficult due to the insensitivity of viral culture, and reverse transcription-PCR (RT-PCR) is a more sensitive alternative. The relationship of quantitative RT-PCR to viable virus has never been studied for RSV. Therefore, we compared a quantitative real-time RT-PCR with viral culture to assess viral load in adult volunteers challenged with the RSV A2 strain. Twelve of 13 volunteers were infected, and there was a high correlation (r = 0.84) between quantitative RT-PCR and viral titer by cell culture. However, RT- PCR was more sensitive, with 73 of 169 (43%) samples positive compared to 58 (34%) samples positive by culture. The correlation between the two tests was highest early in the course of viral shedding (r = 0.91, days 0 to 6), whereas during days 7 to 13, there was more variability (r = 0.70). All subjects were culture negative by day 11, whereas one subject remained RT-PCR positive on day 12. All subjects were RT-PCR negative at day 28 postinfection. Quantitative RT-PCR has an excellent correlation with viral titers, as measured by culture, and should be a useful tool for future studies addressing viral load and disease pathogenesis. Respiratory syncytial virus (RSV) has recently been recognized as a serious pathogen in elderly and immunocompromised adults. Diagnosis of acute infection in adults is often difficult due to the insensitivity of viral culture, and reverse transcription-PCR (RT-PCR) is a more sensitive alternative. The relationship of quantitative RT-PCR to viable virus has never been studied for RSV. Therefore, we compared a quantitative real-time RT-PCR with viral culture to assess viral load in adult volunteers challenged with the RSV A2 strain. Twelve of 13 volunteers were infected, and there was a high correlation ( r = 0.84) between quantitative RT-PCR and viral titer by cell culture. However, RT-PCR was more sensitive, with 73 of 169 (43%) samples positive compared to 58 (34%) samples positive by culture. The correlation between the two tests was highest early in the course of viral shedding ( r = 0.91, days 0 to 6), whereas during days 7 to 13, there was more variability ( r = 0.70). All subjects were culture negative by day 11, whereas one subject remained RT-PCR positive on day 12. All subjects were RT-PCR negative at day 28 postinfection. Quantitative RT-PCR has an excellent correlation with viral titers, as measured by culture, and should be a useful tool for future studies addressing viral load and disease pathogenesis. Respiratory syncytial virus (RSV) has recently been recognized as a serious pathogen in elderly and immunocompromised adults. Diagnosis of acute infection in adults is often difficult due to the insensitivity of viral culture, and reverse transcription-PCR (RT-PCR) is a more sensitive alternative. The relationship of quantitative RT-PCR to viable virus has never been studied for RSV. Therefore, we compared a quantitative real-time RT-PCR with viral culture to assess viral load in adult volunteers challenged with the RSV A2 strain. Twelve of 13 volunteers were infected, and there was a high correlation (r = 0.84) between quantitative RT-PCR and viral titer by cell culture. However, RT-PCR was more sensitive, with 73 of 169 (43%) samples positive compared to 58 (34%) samples positive by culture. The correlation between the two tests was highest early in the course of viral shedding (r = 0.91, days 0 to 6), whereas during days 7 to 13, there was more variability (r = 0.70). All subjects were culture negative by day 11, whereas one subject remained RT-PCR positive on day 12. All subjects were RT-PCR negative at day 28 postinfection. Quantitative RT-PCR has an excellent correlation with viral titers, as measured by culture, and should be a useful tool for future studies addressing viral load and disease pathogenesis.Respiratory syncytial virus (RSV) has recently been recognized as a serious pathogen in elderly and immunocompromised adults. Diagnosis of acute infection in adults is often difficult due to the insensitivity of viral culture, and reverse transcription-PCR (RT-PCR) is a more sensitive alternative. The relationship of quantitative RT-PCR to viable virus has never been studied for RSV. Therefore, we compared a quantitative real-time RT-PCR with viral culture to assess viral load in adult volunteers challenged with the RSV A2 strain. Twelve of 13 volunteers were infected, and there was a high correlation (r = 0.84) between quantitative RT-PCR and viral titer by cell culture. However, RT-PCR was more sensitive, with 73 of 169 (43%) samples positive compared to 58 (34%) samples positive by culture. The correlation between the two tests was highest early in the course of viral shedding (r = 0.91, days 0 to 6), whereas during days 7 to 13, there was more variability (r = 0.70). All subjects were culture negative by day 11, whereas one subject remained RT-PCR positive on day 12. All subjects were RT-PCR negative at day 28 postinfection. Quantitative RT-PCR has an excellent correlation with viral titers, as measured by culture, and should be a useful tool for future studies addressing viral load and disease pathogenesis. |
| Author | Ann R. Falsey John J. Treanor Edward E. Walsh Maria A. Formica |
| AuthorAffiliation | Department of Medicine, 1 Infectious Disease Unit, Rochester General Hospital, 3 Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14621 2 |
| AuthorAffiliation_xml | – name: Department of Medicine, 1 Infectious Disease Unit, Rochester General Hospital, 3 Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14621 2 |
| Author_xml | – sequence: 1 givenname: Ann R. surname: Falsey fullname: Falsey, Ann R. organization: Department of Medicine, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14621 – sequence: 2 givenname: Maria A. surname: Formica fullname: Formica, Maria A. organization: Department of Medicine, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14621 – sequence: 3 givenname: John J. surname: Treanor fullname: Treanor, John J. organization: Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14621 – sequence: 4 givenname: Edward E. surname: Walsh fullname: Walsh, Edward E. organization: Department of Medicine, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York 14621 |
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| Keywords | Virus Human Human respiratory syncytial virus Mononegavirales Pneumovirinae Pneumovirus Viral load Reverse transcription polymerase chain reaction Paramyxoviridae |
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| Notes | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 Corresponding author. Mailing address: Infectious Diseases Unit, Rochester General Hospital, 1425 Portland Ave., Rochester, NY 14621. Phone: (585) 922-4339. Fax: (585) 922-5168. E-mail: ann.falsey@viahealth.org. |
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| SubjectTerms | Adult Antibodies, Viral - analysis Biological and medical sciences Fundamental and applied biological sciences. Psychology Humans Immunoglobulin A, Secretory - analysis Microbiology Middle Aged Nasal Mucosa - virology Respiratory Syncytial Virus, Human - genetics Respiratory Syncytial Virus, Human - isolation & purification Reverse Transcriptase Polymerase Chain Reaction - methods RNA, Viral - analysis Techniques used in virology Viral Load Virology Virus Cultivation Virus Shedding |
| Title | Comparison of Quantitative Reverse Transcription-PCR to Viral Culture for Assessment of Respiratory Syncytial Virus Shedding |
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