MAB8510 Sigma-AldrichAnti-Human Metapneumovirus 75.1 Antibody, clone 1B7

Clinical laboratories historically diagnose seven or eight respiratory virus infections using a combination of techniques including enzyme immunoassay, direct fluorescent antibody staining, cell culture, and nucleic acid amplification tests. With the discovery of six new respiratory viruses since 2000, laboratories are faced with the challenge of detecting up to 19 different viruses that cause acute respiratory disease of both the upper and lower respiratory tracts. The application of nucleic acid amplification technology, particularly multiplex PCR coupled with fluidic or fixed microarrays, provides an important new approach for the detection of multiple respiratory viruses in a single test. These multiplex amplification tests provide a sensitive and comprehensive approach for the diagnosis of respiratory tract infections in individual hospitalized patients and the identification of the etiological agent in outbreaks of respiratory tract infection in the community. This review describes the molecular methods used to detect respiratory viruses and discusses the contribution that molecular testing, especially multiplex PCR, has made to our ability to detect respiratory viruses and to increase our understanding of the roles of various viral agents in acute respiratory disease.

We report a prospective study concerning the efficacy of LLC-MK2 (continuous monkey kidney cell), Hep-2, MDCK (Madin-Darby Canine Kidney), Vero and MRC-5 cell lines, by shell vial assay, and incubation time in the isolation of hMPV from pediatric respiratory samples. The overall sensitivity of the cell lines studied were: 100% for the LLC-MK2, 68.7% for the Hep-2, 28.1% for the Vero, 3.1% for the MDCK and 0% for the MRC-5. Only one strain (3.1%) showed growth in the four cell lines studied and 10 (31.2%) strains only grew in the LLC-MK2 cell line. The analysis of incubation times showed that only 14 strains (43.7%) were able to grow after 3 days of incubation, while all strains (100%) showed growth after 5 days. The use of shell vials with commercial LLC-MK2 cells could be a method for isolating hMPV from respiratory samples in the pediatric population.

Monoclonal antibody MAb-8 was evaluated for detection of human metapneumovirus (HMPV) in shell vial centrifugation cultures (SVCC). Detection of HMPV was similar in A549, HEp-2, and LLC-MK2 SVCC, and MAb-8 staining was optimal on day 2 postinoculation. Availability of SVCC for HMPV will be of significant benefit to clinical laboratories.