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Characterisation of viruses causing human respiratory infections via genomic identification for in vitro diagnosis. Clinical arrays/Clart PneumoVir

Abstract number: P1604

Aleman A., Marcos M.A., Pena M.J., Muir P., Vipond B., Macias A., Villahermosa M.L.

Acute respiratory infections are among the most frequent diseases in our area, and they are one of the main reasons for consultation and hospitalisation. Due to a great variety of possible pathogenic agents and the high frequency of co-infections, it is necessary to use diagnostic methods that allow multiple, sensitive, and rapid identification of these viruses.

Objective: To evaluate the capacity of CLINICAL ARRAYS/CLART®PneumoVir (CA-PV) for simultaneous detection of the 18 most frequent types of human viruses causing respiratory infections:influenza A H1-N1 virus (Flu-A, H1-N1), Flu A H3-N2, Flu-B, Flu-C, parainfluenza virus 1 (PIV-1), PIV-2, PIV-3, PIV-4A, PIV-4B, respiratory syncytial virus type A (RSV-A), RSV-B, human metapneumovirus A (hMPV-A), hMPV-B, rhinovirus, enterovirus, coronavirus, adenovirus, and bocavirus. Several Hospitals from Spain and UK have taken part in this multicentre study.

Method: Virus detection is performed via multiplex RT-PCR for amplification, and a new technology platform based on low-density micro-arrays (ArrayTube) for visualisation. This technology allows simultaneous detection of viruses and any necessary control in order to guarantee the reliability of the results obtained. In order to determine the diagnostic parameters of the kit, a comparative evaluation of CA-PV against clinical diagnostic's current methods (nested-multiplex PCR/agarose gel, culture) was made. A total of 374 clinical specimens were tested, being a true positive result judged according to the concordance between both methods. All the discrepancies were validated with sequencing, and homemade nested-PCR.

Results: 100% analytical sensitivity was obtained in the detection of 14 recombinant plasmids between 10 and 1000 copies. It was observed that there were no cases of unspecific detection of viruses obtaining 100% analytical specificity. About the diagnostic sensitivity and specificity, the behaviour of each virus after the validation of 374 clinical specimens showed that most of virus has sensitivity higher than 90%, and specificity higher than 98%.

Conclusion: CA-PV is useful in the clinical setting for rapid screening and detection of a panel of respiratory viral pathogens based on the following facts: (i) excellent specificities and sensitivities; (ii) rapid and automatic procedure; (iii) simultaneous detection allowing the recognition of co-infections, (iv) hMPV-A, hMPV-B, Flu-A H1N1, Flu-A H3N2, PIV-4A, and PIV-4B subtypes can be differentially detected.

Session Details

Date: 19/04/2008
Time: 00:00-00:00
Session name: 18th European Congress of Clinical Microbiology and Infectious Diseases
Subject:
Location: Barcelona, Spain
Presentation type:
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