Evaluation of direct immunofluorescence, dot-blot enzyme immunoassay, and shell-vial culture for detection of respiratory syncytial virus in patients with bronchiolitis

Reina, Jordi; Ros, M J; Valle, J. M. del; Blanco, Isabel; Munar, María

doi:10.1007/bf01691389

Cited by 10 publications

(12 citation statements)

References 6 publications

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“…The ColorPAC assay was positive and the shell vial culture was negative in 83 (5%) samples; these were initially considered to be probable false‐positives of the antigen detection system. This percentage was lower than that observed (7.7%) in our previous study [14], but was similar to that reported (4.8%) by Ribes et al. [20].…”

Section: Discussionsupporting

confidence: 88%

Prospective evaluation of a dot-blot enzyme immunoassay (Directigen RSV) for the antigenic detection of respiratory syncytial virus from nasopharyngeal aspirates of paediatric patients

Reina

Gárdenas

Gopegui

et al. 2004

Clinical Microbiology and Infection

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This study investigated the efficacy of a commercial enzyme immunoassay (Directigen RSV, ColorPAC) in comparison with the shell vial culture method (using Hep-2 cells) for the detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates from children with bronchiolitis. During the period 1995-2002, 4950 samples were examined. RSV was detected in 1660 (33.5%) samples, with a sensitivity of 80.9%, a specificity of 97.5%, a positive predictive value of 93.8%, a negative predictive value of 91.6%, and a testing efficiency value of 92.2% compared with shell vial culture. In 83 (5%) samples, the ColorPAC was positive and the shell vial assay was negative. Of these, 71 (85.6%) were false-negative by cell culture. The true false-positive results obtained by ColorPAC represented only 0.7% of all RSV-positive samples. In general, no statistically significant differences were detected between the different months and epidemic periods studied. Compared with ColorPAC, the shell vial culture method displayed a sensitivity of 95.8% and a specificity of 100%. Overall, the ColorPAC assay was an acceptable, simple and rapid method for the antigenic detection of RSV in paediatric respiratory samples.

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Section: Discussionsupporting

confidence: 88%

Prospective evaluation of a dot-blot enzyme immunoassay (Directigen RSV) for the antigenic detection of respiratory syncytial virus from nasopharyngeal aspirates of paediatric patients

Reina

Gárdenas

Gopegui

et al. 2004

Clinical Microbiology and Infection

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“…DFA was performed by standard laboratory methods (16,21,22). Briefly, substrate slides were prepared from epithelial cells contained within the NPA specimen, dried, and fixed in acetone for 10 min.…”

Section: Specificity Of Lc-rt-pcrmentioning

confidence: 99%

“…Compared to cell culture and reverse transcriptase PCR (RT-PCR), DFA lacks sensitivity, is subjective, and is dependent on the quality and presence of appropriately infected cells (30). Although cell culture is more sensitive than DFA, it requires specimens to be transported and stored under ideal conditions, and the prolonged turnaround time required to obtain results further diminishes its usefulness in patient management (22).…”

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confidence: 99%

Detection of Human Respiratory Syncytial Virus in Respiratory Samples by LightCycler Reverse Transcriptase PCR

et al. 2002

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“…DFA detection is more rapid but less sensitive than viral culture, and results may be affected by specimen quality (ie, presence of intact, infected cells), virus type, and interpretation of a positive result, which is subjective and requires a great deal of technical skill. 2,[7][8][9][10] DFA is also unable to detect minor variations in amino acid sequence on envelope or capsid proteins. 11 Viral culture is still considered the "gold standard" for respiratory virus detection, but is limited by a prolonged result turnaround time (ie, 2 days to 1 week) and is dependent on stringent specimen transport and storage conditions to preserve the infectivity of the virus.…”

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confidence: 99%

A Sensitive, Specific, and Cost-Effective Multiplex Reverse Transcriptase-PCR Assay for the Detection of Seven Common Respiratory Viruses in Respiratory Samples

Syrmis¹,

Whiley²,

Thomas³

et al. 2004

The Journal of Molecular Diagnostics

159

153

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Common etiological viral agents of respiratory infections include adenoviruses (ADV), influenza types A and B (Flu A and B), parainfluenza types 1, 2, and 3 (PIV1, 2, 3),and respiratory syncytial virus (RSV). 1-4 These viruses are responsible for a spectrum of acute upper and lower respiratory tract disease. In children, the elderly and other immunocompromised groups, respiratory viruses can cause more serious clinical complications, such as croup, bronchiolitis, and pneumonia, which often require hospitalization. 5,6 Virus isolation by cell culture and direct immunofluorescent antibody assay (DFA) staining with monoclonal antibodies are two of the most commonly used laboratory techniques for detecting respiratory viruses. Both these methods have significant limitations in sensitivity and specificity. DFA detection is more rapid but less sensitive than viral culture, and results may be affected by specimen quality (ie, presence of intact, infected cells), virus type, and interpretation of a positive result, which is subjective and requires a great deal of technical skill. 2,7-10 DFA is also unable to detect minor variations in amino acid sequence on envelope or capsid proteins. 11 Viral culture is still considered the "gold standard" for respiratory virus detection, but is limited by a prolonged result turnaround time (ie, 2 days to 1 week) and is dependent on stringent specimen transport and storage conditions to preserve the infectivity of the virus. 9,[12][13][14][15] Although the combination of both these techniques can provide an increase in the number of positive results, a significant proportion of specimens still remain negative, despite clinical suspicion of viral infection. 8 Several studies have shown that polymerase chain reaction (PCR) amplification can resolve the intrinsic limitations associated with traditional diagnostic techniques by combining increased sensitivity, specificity, and rapid result turnaround time. 16,17 Also, PCR results are not dependent on infectious virus or viable cells. However, Supported by Royal Children's Hospital Foundation grants RA921-006 and I922-034 which were sponsored by the Woolworth's "Care for Kids" campaign.

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Evaluation of direct immunofluorescence, dot-blot enzyme immunoassay, and shell-vial culture for detection of respiratory syncytial virus in patients with bronchiolitis

Cited by 10 publications

References 6 publications

Prospective evaluation of a dot-blot enzyme immunoassay (Directigen RSV) for the antigenic detection of respiratory syncytial virus from nasopharyngeal aspirates of paediatric patients

Prospective evaluation of a dot-blot enzyme immunoassay (Directigen RSV) for the antigenic detection of respiratory syncytial virus from nasopharyngeal aspirates of paediatric patients

Detection of Human Respiratory Syncytial Virus in Respiratory Samples by LightCycler Reverse Transcriptase PCR

A Sensitive, Specific, and Cost-Effective Multiplex Reverse Transcriptase-PCR Assay for the Detection of Seven Common Respiratory Viruses in Respiratory Samples

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