Microarray-based genotyping of Salmonella: Inter-laboratory evaluation of reproducibility and standardization potential

摘要: Bacterial food-borne infections in humans caused by Salmonella spp. are considered a crucial food safety issue. Therefore, it is important for the risk assessments of to consider genomic variation among different isolates order control pathogen-induced infections. Microarray technology promising diagnostic tool that provides information on many genes simultaneously. However, standardization DNA microarray analysis needed before can be used as routine method characterizing across borders and laboratories. A comparative study was designed which agreement data from assay typing between two labs assessed. The expected reveal possibility obtaining same results using equipment evaluate reproducibility technique first step towards standardization. low-density array contains 281 57-60-mer oligonucleotide probes detecting wide range specific marker associated with antibiotic resistance, cell envelope structures, mobile genetic elements pathogenicity. Several critical methodology parameters differed were identified. These related printing facilities, choice hybridization buffer, wash buffers following procedure purifying DNA. Critical randomized four-factorial experiment statistical measures inter-lab consistency performed based kappa coefficient. high level (kappa=0.7-1.0) obtained even when employing procedures buffers. less (Kappa=0.2-0.6) observed buffers, indicating this parameter being highly transferring standard In conclusion, indicates assays reproduced at least pre-requisite development guidelines.