Mkangara Mwanaisha, Mbega Ernest R, Chacha Musa
Department of Sustainable Agriculture and Biodiversity and Ecosystems Management, Nelson Mandela African Institution of Science and Technology, Arusha, Tanzania.
Department of Science and Laboratory Technology, Dar es Salaam Institute of Technology, Dar es Salaam, Tanzania.
Vet World. 2019 Apr;13(4):764-767. doi: 10.14202/vetworld.2020.764-767. Epub 2020 Apr 23.
This study aimed to identify serovars by polymerase chain reaction (PCR) based on virulence genes invasion A () and plasmid virulence C ().
DNA extraction of eight bacteria isolates was done using the PowerSoil DNA Isolation Kit. The amplification of and genes was done using conventional PCR. The positive PCR products were purified using the GeneJET Purification Kit and then sequenced using ABI 3730 XL automated genetic analyzer. The sequences obtained were compared for similarities with other serovars deposited on the NCBI GenBank using BLASTN.
Four out of eight samples were amplified by primers FS139/RS141 that target gene with products of about 284 bp, and three out of four of the same positive samples were also amplified by primers FSPV-1/RSPV-2 targeting with a product of about 571 bp. One sample was not amplified by primers FSPV-1/RSPV-2 as it lacked virulence plasmid. Analysis of sequences indicated 100% homology with closely related serovars of . subspecies enterica serovar Typhimurium.
Typhimurium that contained and genes are pathogenic and virulent strains.
本研究旨在通过基于毒力基因侵袭A（）和质粒毒力C（）的聚合酶链反应（PCR）来鉴定血清型。
使用PowerSoil DNA提取试剂盒对八株细菌分离株进行DNA提取。使用常规PCR对和基因进行扩增。使用GeneJET纯化试剂盒对PCR阳性产物进行纯化，然后使用ABI 3730 XL自动基因分析仪进行测序。使用BLASTN将获得的序列与NCBI GenBank上保存的其他血清型进行相似性比较。
八份样本中有四份被靶向基因的引物FS139/RS141扩增，产物约为284 bp，在这四份相同的阳性样本中，有三份也被靶向的引物FSPV-1/RSPV-2扩增，产物约为571 bp。有一份样本未被引物FSPV-1/RSPV-2扩增，因为它缺乏毒力质粒。序列分析表明与肠炎沙门氏菌亚种鼠伤寒血清型的密切相关血清型具有100%的同源性。
含有和基因的鼠伤寒沙门氏菌是致病的有毒力菌株。
