Evaluation comparative de huit trousses commerciales de PCR multiplex pour le diagnostic des infections respiratoires

LYON1-BU Santé (693882101) / SudocSudocFranceF

Comparative evaluation of six commercialized multiplex PCR kits for the diagnosis of respiratory infections.

The molecular diagnosis of respiratory infection can be performed using different commercial multiplex-based PCR kits whose performances have been previously compared individually to those of conventional techniques. This study compared the practicability and the diagnostic performances of six CE-marked kits available in 2011 on the French market, including 2 detecting viruses and atypical bacteria (from Pathofinder and Seegene companies) and 4 detecting only viruses (from Abbott, Genomica, Qiagen and Seegene companies). The respective sensitivity, specificity, accuracy and agreement of each multiplex technique were calculated by comparison to commercial duplex PCR tests (Argene/bioMérieux) used as gold standard. Eighty-eight respiratory specimens with no pathogen (n = 11), single infections (n = 33) or co-infections (n = 44) were selected to cover 9 viruses or groups of viruses and 3 atypical bacteria. All samples were extracted using the NUCLISENS® easyMAG™ instrument (bioMérieux). The overall sensitivity ranged from 56.25% to 91.67% for viruses and was below 50% with both tests for bacteria. The overall specificity was excellent (>94% for all pathogens). For each tested kit, the overall agreement with the reference test was strong for viruses (kappa test >0.60) and moderate for bacteria. After the extraction step, the hands-on time varied from 50 min to 2h30 and the complete results were available in 2h30 to 9 h. The spectrum of tested agents and the technology used to reveal the PCR products as well as the laboratory organization are determinant for the selection of a kit

Presentation of the 7 kits used in the study for detecting viruses (see text for correspondence between letters and commercial denominations).

<p>Yes: group of pathogens detected; w diff: with differentiation between types. No: group of pathogens not detected.</p

List of pathogens that were considered to be present in the panel of the 88 specimens according the combination of duplex PCR tests (Argene/bioMérieux) used as gold standard.

a<p>type 1 (n = 2), type 3 (n = 3) and type 4 (n = 5).</p>b<p>type NL63 (n = 6), type OC43 (n = 8), type 229E (n = 1), type HKU1 (n = 6) and untyped (n = 4).</p

Practicability of each kit using the NUCLISENS easyMAG as extraction instrument (see text for correspondence between letters and commercial denominations).

<p>Practicability of each kit using the NUCLISENS easyMAG as extraction instrument (see text for correspondence between letters and commercial denominations).</p

Performances of the 6 kits evaluated in this study for a panel of 9 viruses or groups of viruses with reference to the combination of duplex PCR tests (Argene/bioMérieux) used as gold standard.

a<p>Kit having no probe for the detection of HKU1 coronavirus.</p>b<p>Kit having a probe for detecting 229E coronavirus only.</p>c<p>All the tested kits did not detect all the coronavirus types (see notes ^a and ^b).</p

Schematic synopsis of the design of the study.

<p>(See text for correspondence between letters and commercial denominations).</p