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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
Author(s) -
Khan Shams Tabrez,
Ahmad Javed,
Wahab Rizwan,
Hirad Abdurahman H.,
Musarrat Javed,
AlKhedhairy Abdulaziz A.,
Bahkali Ali H.
Publication year - 2018
Publication title -
journal of food safety
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 43
eISSN - 1745-4565
pISSN - 0149-6085
DOI - 10.1111/jfs.12381
Subject(s) - raw milk , brucella , brucellosis , 16s ribosomal rna , biology , genomic dna , population , polymerase chain reaction , camel milk , veterinary medicine , primer (cosmetics) , dna extraction , gene , microbiology and biotechnology , food science , medicine , genetics , virology , chemistry , environmental health , organic chemistry
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, omp 2, and IS 711 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 applications This 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.