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Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopathological assessment.

ABSTRACT: BACKGROUND:Infections with common respiratory tract viruses can cause high mortality, especially in immunocompromised hosts, but the impact of human metapneumovirus (hMPV) in this setting was previously unknown. METHODS:We evaluated consecutive bronchoalveolar lavage and bronchial wash fluid samples from 688 patients--72% were immunocompromised and were predominantly lung transplant recipients--for hMPV by use of quantitative real-time polymerase chain reaction (PCR), and positive results were correlated with clinical outcome and results of viral cultures, in situ hybridization, and lung histopathological assessment. RESULTS:Six cases of hMPV infection were identified, and they had a similar frequency and occurred in a similar age range as other paramyxoviral infections. Four of 6 infections occurred in immunocompromised patients. Infection was confirmed by in situ hybridization for the viral nucleocapsid gene. Histopathological assessment of lung tissue samples showed acute and organizing injury, and smudge cell formation was distinct from findings in infections with other paramyxoviruses. Each patient with high titers of hMPV exhibited a complicated clinical course requiring prolonged hospitalization. CONCLUSIONS:Our results provide in situ evidence of hMPV infection in humans and suggest that hMPV is a cause of clinically severe lower respiratory tract infection that can be detected during bronchoscopy by use of real-time PCR and routine histopathological assessment.

SUBMITTER: Sumino KC

PROVIDER: S-EPMC7202407 | biostudies-literature | 2005 Sep

REPOSITORIES: biostudies-literature

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Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopathological assessment.

Sumino Kaharu C KC Agapov Eugene E Pierce Richard A RA Trulock Elbert P EP Pfeifer John D JD Ritter Jon H JH Gaudreault-Keener Monique M Storch Gregory A GA Holtzman Michael J MJ

The Journal of infectious diseases 20050804 6

<h4>Background</h4>Infections with common respiratory tract viruses can cause high mortality, especially in immunocompromised hosts, but the impact of human metapneumovirus (hMPV) in this setting was previously unknown.<h4>Methods</h4>We evaluated consecutive bronchoalveolar lavage and bronchial wash fluid samples from 688 patients--72% were immunocompromised and were predominantly lung transplant recipients--for hMPV by use of quantitative real-time polymerase chain reaction (PCR), and positive ...[more]

PMID: 16107959

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Project description:The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5' untranslated region (5' UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.

Dataset Information

Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopathological assessment.

Publications

Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopathological assessment.

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