Rapid Laboratory Diagnostics during the Winter Respiratory Virus Season

Lipson, Steven M.

doi:10.1128/jcm.40.2.733-734.2002

Cited by 8 publications

(6 citation statements)

References 13 publications

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“…We have described the detection of hMPV from Australian children and infants by two newly developed RT-PCR assays. The need for rapid diagnosis of respiratory tract disease is acutely important during seasonal outbreaks of the causative agents (6).…”

Section: Discussionmentioning

confidence: 99%

Molecular Assays for Detection of Human Metapneumovirus

et al. 2003

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The recent description of the respiratory pathogen human metapneumovirus (hMPV) has highlighted a deficiency in current diagnostic techniques for viral agents associated with acute lower respiratory tract infections. We describe two novel approaches to the detection of viral RNA by use of reverse transcriptase PCR (RT-PCR). The PCR products were identified after capture onto a solid-phase medium by hybridization with a sequence-specific, biotinylated oligonucleotide probe. The assay was applied to the screening of 329 nasopharyngeal aspirates sampled from patients suffering from respiratory tract disease. These samples were negative for other common microbial causes of respiratory tract disease. We were able to detect hMPV sequences in 32 (9.7%) samples collected from Australian patients during 2001. To further reduce result turnaround times we designed a fluorogenic TaqMan oligoprobe and combined it with the existing primers for use on the LightCycler platform. The real-time RT-PCR proved to be highly reproducible and detected hMPV in an additional 6 out of 62 samples (9.6%) tested during the comparison of the two diagnostic approaches. We found the real-time RT-PCR to be the test of choice for future investigation of samples for hMPV due to its speed, reproducibility, specificity, and sensitivity.

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Section: Discussionmentioning

confidence: 99%

Molecular Assays for Detection of Human Metapneumovirus

et al. 2003

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“…Real-time platforms also offer rapid turnaround times with the ability to test for a number of gene targets with multiplexing. With PCR leading the way for the detection of viral agents [Lipson, 2002], we have developed and validated two real-time assays targeting the non-structural protein (NP-1) gene and the viral protein (VP-1) gene for the detection of HBoV in three different sample types, combined nose-throat swabs (NTS), fecal samples, and whole blood.…”

Section: Introductionmentioning

confidence: 99%

Detection of human bocavirus in respiratory, fecal, and blood samples by real‐time PCR

Tozer

Lambert

Whiley

et al. 2009

Journal of Medical Virology

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Human bocavirus (HBoV) has been detected worldwide in respiratory samples. Two real-time PCR assays, targeting the non-structural protein (NP-1) and viral protein (VP-1) genes, were designed and validated to detect HBoV in patients with respiratory disease, gastroenteritis, or systemic illness. Sensitivity of the NP-1 and VP-1 assays were equal to the conventional PCR assay previously described by Allander et al. [2005: Proc Natl Acad Sci USA 102: 12891-12896] being 100%, and giving specificity of 94% and 93%, respectively. There was no cross-reaction identified with unrelated respiratory agents, or to human DNA. The limits of detection were 10 copies of genomic DNA equivalents per reaction for both assays. The assays were used to screen three different sample populations, combined nose, and throat swabs (n = 96) from children with acute respiratory disease, fecal samples (n = 375) from adults, and children with gastroenteritis and whole blood (n = 229) collected from 31 immunocompromised children taken over an 18-month period. In total 17 (18%) respiratory samples and 18 (4.8%) fecal samples were identified as having HBoV present. Of the pediatric whole blood specimens investigated, HBoV was detected in six (2.6%) samples from four patients. In summary, two real-time PCR assays targeting different genes were designed and validated for use as screening methods for the detection of HBoV. HBoV was found in three different specimen types: parent-collected combined nose-throat swabs, fecal samples collected from symptomatic individuals and whole blood from immunocompromised children.

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“…We summarized these for convenience because the evaluation of singular tests is not our main focus. Antigen tests have the advantage of quickly yielding results [9,10], whereas PCR and isolation analyses require a specialized laboratory, and results are thus not readily available. The evaluation of antigen tests with virus isolation as the RS revealed sensitivities and specificities that are considered to be high but not one [8].…”

Section: Diagnostic Testsmentioning

confidence: 99%