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J. Clin. Microbiol. doi:10.1128/JCM.01567-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Clinical Evaluation of NucliSENS Magnetic Extraction and NucliSENS Analytic Specific Reagents for the Real Time Detection of Human Metapneumovirus (hMPV) in Pediatric Respiratory Specimens

North Shore – Long Island Jewish Health System Laboratories, Department of Laboratory Medicine, Lake Success, NY, North Shore University Hospital, Department of Laboratory Medicine, Manhasset, NY; North Shore – Long Island Jewish Health System Laboratories, Molecular Diagnostics, Department of Laboratory Medicine, 10 Nevada Drive, Lake Success, NY 11042; North Shore University Hospital, Clinical Virology, Department of Laboratory Medicine, 300 Community Drive, Manhasset, NY 11030

^* To whom correspondence should be addressed. Email: cginocch{at}nshs.edu.

	Abstract

In this study, we evaluated the NucliSENS miniMAG (MM) and easyMAG (EM) nucleic acid extraction platforms (bioMerieux, Durham, NC) in combination with the NucliSENS EasyQ Basic kit and analyte specific reagents (bioMérieux) for the detection of human metapneumovirus (hMPV) in respiratory samples. Total nucleic acids from pediatric clinical samples (n=653) and an hMPV specific inhibition control (h-IC) were co-extracted using the MM and/or the EM. Nucleic acid sequence based amplification (NASBA) and real time molecular beacon detection of hMPV was performed using a NucliSENS EasyQ Analyzer (bioMérieux). Positive results were confirmed using an in-house validated RT-PCR ASR based assay. The inclusion of the h-IC monitored the entire process, including the efficiency of nucleic acid extraction, amplification and detection. The percentage of samples with inhibited amplification of the h-IC after initial NA extraction by EM and MM were 1.88% and 3.17%, respectively. After reprocessing of a new aliquot, the final h-IC inhibition rates were 0% (EM) and 1.06% (MM). The limit of detection of the assay was between 2 (EM extraction) and 10 (MM extraction) RNA copies/reaction and specificity was 100% when testing viral respiratory isolates and clinical samples. hMPV was detected in 5.6% of pediatric samples tested and was detected in 3 co-infections with respiratory syncytial virus (RSV). hMPV was the second most frequently detected respiratory virus in children 0-2 yrs of age, after RSV. In summary, NucliSENS extraction and ASRs provided a sensitive and specific method for the detection of hMPV in respiratory samples.