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在MAGPIX上运行xTAG多重肠胃道病原体检测试剂盒的初步评估

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发布日期:2011-11-04 16:56 文章来源:路明克斯
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Background: The xTAG® Gastrointestinal Pathogen Panel (GPP) from Luminex Molecular Diagnostics (Toronto, Canada) is a new qualitative nucleic acid multiplex assay designed for the simultaneous detection of 15 pathogens in human stool samples,including 3 viruses,3 parasites and 9 bacteria/bacteria toxins. The assay is run on either the Luminex® 100/200TM or the recently introduced, MAGPIX instrument that utilizes the CCD imaging technology. In this study, we examine the performance of the GPP test using the MAGPIX platform for the detection of 4 targets: Adenovirus (serotypes 40 and 41), Norovirus (GI /GII), Rotavirus A and C. difficile (Toxin A/B).

 

Methods:  One hundred and fourteen stool specimens were selected for evaluation including 26 Rotavirus, 33 Norovirus, 15 Adenovirus, and 34 C. difficile positives.  Nucleic acid was extracted from 140 μl of 10% stool suspensions using the bioMerieux easy MAGTM extractor. GPP testingn was performed according to maufacturer's instructions. Briefly, reverse transcription and amplification was performed in a one-step reaction, amplified products were sorted on a bead-array incorporating the xTAG Universal Array coupled to magnetic microspheres, and reactions read on the MAGPIX instrument. Results were compared to reference methods, including the RIDA Quick Rotavirus (R-Biopharm) for Rotavirus and in-house developed PCR assays for Norovirus, Adenovirus and C. difficile.
 

Results: Of 114 stools tested, the GPP assay detected 37/37 Norovirus positives (2 GI and 35 GII genotypes), 26/26 Rotavirus A positives and 5/5 Adenovirus serotypes 40/41. Ten Adenovirus positive stools that were confirmed to be non-40/41 serotypes by sequencing or genotype-specific PCR were negative when tested by the GPP assay as expected.  For C. difficile, GPP detected 34/37 (91.9%) positive stools.  Of the 3 C. difficile specimens that were missed by GPP, crossing
points for the in-house assay were greater than 36 cycles, indicating low level positives. The GPP detected 7/114 (6.1%) stools positive for more than one pathogen including 1 Rotavirus/ C. difficile, 2 Norovirus/ C. difficile, 1 Adenovirus/ C. difficile, 1 Norovirus/ Rotavirus and 2 Salmonella sp. / C. difficile
 

Conclusions:  In this preliminary evaluation, the xTAG Gastrointestinal Pathogen Panel had excellent sensitivity and specificity.  The overall sensitivity and specificity for these four targets was 97.4% (111/114) and 100% respectively.  The assay was run on the less expensive MAGPIX instrument but can also be run on the Luminex 100/200.  Further investigations are required to examine the performance of the other pathogens.
 

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