An improved method of DNA preparation for PCR‐based detection of Brucella in raw camel milk samples from Riyadh region and its comparison with immunological methods
JOURNAL OF FOOD SAFETY(2018)
摘要
Brucellosis a zoonotic disease is endemic in Saudi Arabia especially in the Najed region due to the customary consumption of raw milk. For a detailed investigation of brucellosis in camel milk, 80 samples were collected from Riyadh region. Twenty-four and sixteen percentage of the samples were positive for Brucella when tested with Rose Bengal test and ELISA, respectively. Genomic DNA from the milk samples was prepared using a modified method presented in this study. Brucella was detected in camel milk using three different primer sets targeting Brucella 16S rRNA gene, omp2, and IS711 genes. Among the three primer sets, an F4/R2 set of primers targeting Brucella 16S rRNA gene was most sensitive and yielded PCR products from nine milk samples. Finally, the 16S rRNA gene sequences of these isolates confirmed the contamination of Brucella spp. in nine milk samples. This study, therefore, suggests a high incidence of Brucella in camels of Riyadh. Practical applicationsThis manuscript describes a modified method for the isolation of DNA from camel milk. The DNA was subsequently used successfully for the amplification of various genes for detecting Brucella. Furthermore, there was no report from Riyadh region on the detection of Brucella from camel milk. This report, therefore, points out toward an important health issue associated with consumption of raw camel milk. If published, this report will be helpful in educating the local population about the health risk associated with the consumption of raw milk.
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