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DETECTION OF HUMAN METAPNEUMOVIRUS INFECTION IN CHILDREN ANDADULTS BY MOLECULAR BASED METHODS

Dare, Ryan Keith (2005) DETECTION OF HUMAN METAPNEUMOVIRUS INFECTION IN CHILDREN ANDADULTS BY MOLECULAR BASED METHODS. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

Human metapneumovirus (hMPV) is a recently discovered paramyxovirus known to causerespiratory 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%) andchildren with respiratory symptoms (1.83%). This research is of major public health significancedue to the amount of respiratory infections that are going undiagnosed or being treated withunnecessary antibiotics. It is important for our physicians to not only know that hMPV is presentin 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.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Dare, Ryan Keithryan3321@aol.com
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairRinaldo, Charlesrinaldo@pitt.eduRINALDO
Committee MemberBunker, Clareannbunkerc@pitt.eduBUNKERC
Committee MemberJenkins, Frankfjenkins@pitt.eduFJENKINS
Date: 8 July 2005
Date Type: Completion
Defense Date: 23 May 2005
Approval Date: 8 July 2005
Submission Date: 9 June 2005
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: Graduate School of Public Health > Infectious Diseases and Microbiology
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: real time NASBA; real time RT-PCR
Other ID: http://etd.library.pitt.edu/ETD/available/etd-06092005-091136/, etd-06092005-091136
Date Deposited: 10 Nov 2011 19:46
Last Modified: 15 Nov 2016 13:44
URI: http://d-scholarship.pitt.edu/id/eprint/8049

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