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Whole genome sequence of Brucella melitensisl local isolate from an infected goat in Malaysia
2016
Mohd Mokhtar Arshad | Ramlan Mohamed | Shuhaila Mat Sharani | Hardy Abu Daud | Omer Khazaal Sallou | Mohd Azam Khan Goriman Khan | Hirzahida Mohd. Padil
Brucellosis in goats is mainly caused by the bacterium Brucellamelitensis, which is one of the most important pathogenic species in the world. In Malaysia, the annual prevalence data of brucellosis was recorded in goats and the control strategy of the disease basedon test and cull of infected animals. This strategy has caused huge economic losses to farmers and government alike. Therefore, whole genome sequencing of B. melitensis local strain is essential forimproving the current vaccine. B. melitensis strain VRI 6530/11 wasobtained from veterinary research institute biobank, Ipoh. The strain was submitted for classical identification procedures and the total genomic DNA was extracted by using DNeasy blood and tissue kit(QIAGEN). The concentration and purity of DNA were determined by using agarose gel electrophoresis and spectrophotometer (DNA/RNA) assay respectively. The genome was sequenced by using IlluminaHiSeq platform with insert size ~200 bp. A total of 1.0 Gb data was generated from the sample. More than 95% of sequencing data was retained in the sample after quality filtering, this indicatethe sequencing reads are of high quality. Final assembly had 33 scaffolds with total size ~3.28 Mb, 44 contigs, GC content is 57.25%, N50 is 293,291. A total of 3,238 protein coding genes, 48 tRNAs and 3rRNAs were predicted and over 87% of the genes were functionally annotated. Genome sequencing of a local B. melitensis strain is the first of its kind in Malaysia and work from this study can contribute towards the development of a new effective vaccine for the control ofthe disease in the country.
Afficher plus [+] Moins [-]Development of an in-house Rose Bengal plate test for diagnosis of Brucellosis in goat
2016
Mohamed Ariff O. | Siti Khairani Bejo | Asinamai Athliamai Bitrus | Sani M. Y. | Zakaria Zunita
Brucellosis, caused by Brucella melitensis, is a significantproblem for both public and animal health worldwide. The Rose Bengal plate test (RBPT) antigen from Brucella melitensis local isolates were developed in this study. The performance of the assay wasinvestigated using serum samples collected from goats. A total of 1063 serum samples obtained from goats were examined for thepresence of antibodies against Brucella by in-house RBPT (LRBPT), commercial RBPT (Veterinary Laboratory Agency – VLA, UK) and Compliment Fixation test (CFT). The sensitivity and specificity wascalculated using CFT as the gold standard. Out of 1063 goats sera analyzed 364 (34.24%), 335 (31.51%), and 373 (35.08%) were found to be positive by LRBPT, commercial RBPT and CFT, respectively. The sensitivity calculated for the LRBPT, was 90.1% compared to commercial RBPT 85.0%. However, the specificity of the LRBPT was lower (95.9%), than the commercial RBPT (97.4%). Furthermorethe LRBPT has better value of NPV (94.7%) than commercial RBPT NPV(92.3%). While the PPV, of the commercial RBPT is higher (94.6%) than LRBPT (92.3%) respectively. High sensitive and low cost LRBPT compared to cRBPT B. melitensis RBPT test was successfully developed in this present study. Therefore it was concluded that this diagnostic test kit can complement and replace the availablecommercial RBPT which is relatively more expensive and less sensitive in detection of brucellosis in goats. It could also be used for epidemiological surveillance of caprine brucellosis in Malaysia.
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