Human metapneumovirus (hMPV) is a recently discovered paramyxovirus known to cause
respiratory tract infections primarily in children. This previously unknown pathogen remained undetected for years due to very slow replication in vitro and an inconsistent CPE. More recently, detection of hMPV by means of quantitative molecular techniques has proved to be more effective than culture methods. In this study we describe the development of a quantitative real time RT-PCR assay targeting the hMPV nucleoprotein (N) gene. This assay is compared to a real time nucleic acid sequence based amplification (NASBA) test, developed by bioMérieux, using control material from hMPV strains Can97-83 and Can98-75 representative of the two main lineages A and B, respectively. Using control material the real time RT-PCR, designed to detect all four sublineages of hMPV, can detect as low as 50 and 100 copies of viral RNA from the A and B lineages respectively. The real time NASBA assay can also detect 50 copies of viral RNA from the A strain but only detects 1000 copies of strain B viral RNA. In this study,
hMPV has been detected in both immunosuppressed lung transplant recipients (2.14%) and
children with respiratory symptoms (1.83%). This research is of major public health significance
due to the amount of respiratory infections that are going undiagnosed or being treated with
unnecessary antibiotics. It is important for our physicians to not only know that hMPV is present
in our community but also to be able to detect and treat it appropriately. This study reports the first evidence of hMPV in the Pittsburgh area and demonstrates the importance of this virus as a critical player among respiratory pathogens in both immunosuppressed lung transplant recipients and children. In conclusion, we have successfully developed a real time RT-PCR assay targeting the hMPV N gene. Using this assay along with the real time NASBA assay developed by bioMérieux, we have detected hMPV infections in lung transplant recipients in a year long study. Using the real time RT-PCR assay alone hMPV has also been detected in children suspected of respiratory infection during the early winter season.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-06092005-091136 |
| Date | 08 July 2005 |
| Creators | Dare, Ryan Keith |
| Contributors | Clareann Bunker, Frank Jenkins, Charles Rinaldo |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-06092005-091136/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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