Sunday, October 3, 2010

High-throughput Universal Probe Salmonella Serotyping (UPSS) by nanoPCR


A new publication from the September 2010 Journal of Microbiological Methods discusses the use of nano liter PCR to detect Salmonella. This is quite interesting opportunity in automating and miniaturizing PCR procedures. Here is the abstract:

Salmonella enterica subsp. enterica serovar identification is of great importance with respect to outbreak monitoring and case verification. Therefore rapid, sensitive and cost efficient detection of Salmonella spp is indispensable within microbiology labs. To amalgamate single tube isolate identification with Salmonella typing, we developed the high-throughput Universal Probe Salmonella Serotyping (UPSS) technique based on nano liter PCR. In comparison to the classical approach, where O- and H-antisera are applied, the UPSS relies on specific gene content amplification of Salmonella spp. by an universal TaqMan assay for all markers and identification of the specific amplicon pattern. To enable high-throughput technology we employed a chip format containing 1024 wells loaded by an automated liquid-handling system which allowed us to perform TaqMan PCR reactions in volumes of 100 nano liters per well. Herein we present proof of principle of the UPSS method by the use of a test panel of 100 previously serotyped Salmonella isolates to successfully verify the usability, accuracy and feasibility of the newly developed UPSS approach. We found that the methodology of the UPSS technology is capable of unequivocally identifying 30 Salmonella serotypes on a single chip within 3 hours but can be highly parallelized by the use of multiple PCR machines. Therefore the UPSS method offers a robust and straightforward molecular alternative for Salmonella detection and typing that saves expensive chemistry and can be easily automated.

Authors: Mertes F, Biens K, Lehrach H, Wagner M, and Dahl A. from the Max Planck Institute for Molecular Genetics, Ihnestrasse 73, 14195 Berlin, Germany.

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