Further evaluation and validation of a commercially available competitive ELISA (cELISA) for the detection of antibodies specific to equine arteritis virus (EAV).

The purpose of this study was to further evaluate and validate two commercially available equine arteritis virus (EAV) competitive ELISAs (original and enhanced cELISAs) using archived equine sera from experimentally inoculated animals and field sera submitted for laboratory diagnosis. First, the original and subsequently enhanced cELISAs were compared with the virus neutralisation test (VNT) using a panel of archived serum samples from experimentally inoculated animals. Then, the enhanced cELISA was compared with the VNT using a large panel of archived serum samples. The total number of equine sera tested was 3255, which included sera against 25 different EAV strains. The study confirmed that the enhanced cELISA was more sensitive than the original cELISA. Based on testing sera from experimentally inoculated animals and field sera, the enhanced cELISA had an estimated sensitivity (98.9 percent and 99.6 percent, respectively) and specificity (98.3 percent and 98.7 percent, respectively). The currently marketed enhanced VMRD EAV antibody cELISA test kit (VMRD Inc., Pullman, Washington, USA) has high sensitivity and specificity relative to the VNT. Based on the findings of this study, the authors would propose that the enhanced cELISA should be considered as an alternative approved method to the VNT for the detection of antibodies to EAV.

Performance of the Kallestad Pathfinder enzyme immunoassay in the diagnosis of respiratory syncytial virus infections.

The Kallestad Pathfinder enzyme immunoassay (EIA) for the rapid detection of respiratory syncytial virus (RSV) antigen was compared with virus culture and direct fluorescent antibody (DFA) to determine the reliability of the EIA. During two consecutive winter respiratory seasons, 270 nasopharyngeal wash specimens were tested. RSV was detected in culture by the presence of cytopathic effect and/or an indirect immunofluorescence assay. The sensitivity of the Pathfinder EIA in comparison with isolation in tube culture was 72% (73 of 101) and the specificity was 99% (167 of 169). During the second year of the evaluation period, DFA was performed on all specimens. The sensitivity of the DFA compared with isolation in tube culture was 94%. This study indicates that the Pathfinder EIA is a very specific test for diagnosis of RSV infections, but lacks sensitivity in comparison with tube culture or direct immunofluorescence.

Isolation of seven respiratory viruses in shell vials: a practical and highly sensitive method.

The isolation of respiratory viruses in shell vials was compared with isolation in tube cultures in order to determine the sensitivity of the former, rapid method. Twenty of 21 influenza virus and 15 of 15 parainfluenza virus isolates were recovered in shell vials. One hundred twenty-seven of 138 respiratory syncytial virus isolates were detected in shell vials, but only 10 of 21 adenovirus isolates were positive by the rapid method. Shell vials are very effective for the diagnosis of respiratory viral infections, except for those caused by adenovirus.

Evaluation of Abbott TestPack RSV for the diagnosis of respiratory syncytial virus infections.

Abbott TestPack RSV, a 20-minute enzyme immunoassay, is available for the rapid diagnosis of respiratory syncytial virus (RSV) infections. We have compared TestPack with a "gold standard" method of virus isolation in traditional tube cultures and shell vials to determine the sensitivity and specificity of this rapid method. Respiratory specimens were collected prospectively from 402 children and assayed by the rapid antigen detection method and isolation in culture. Virus was isolated by inoculation of specimen in a total of eight tubes and 2-3 shell vials. Isolation of RSV was confirmed by characteristic cytopathic effect and immunofluorescence using monoclonal antibodies to RSV. Of the 402 specimens tested, there were only 18 discrepant results (seven TestPack-positive, culture-negative, and 11 TestPack-negative, culture-positive specimens). The sensitivity of TestPack RSV versus culture was 93.6% (162 of 173) and the specificity was 97.0% (222 of 229). Using a very rigorous culture system, we have obtained high values for the sensitivity and specificity of TestPack RSV. This assay is an excellent method for the rapid diagnosis of RSV infections in young children.