Molecular identification of Salmonella Typhimurium from village chickens based on invA and spvC genes

Abstract

<jats:p>Aim: This study aimed to identify Salmonella enterica serovars by polymerase chain reaction (PCR) based on virulence genes invasion A (inv A) and Salmonella plasmid virulence C (spvC). Materials and Methods: DNA extraction of eight bacteria isolates was done using the PowerSoil® DNA Isolation Kit. The amplification of invA and spvC 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 Salmonella serovars deposited on the NCBI GenBank using BLASTN. Results: Four out of eight samples were amplified by primers FS139/RS141 that target invA gene with products of about 284 bp, and three out of four of the same invA positive samples were also amplified by primers FSPV-1/RSPV-2 targeting spvC 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 S. enterica subspecies enterica serovar Typhimurium. Conclusion: Salmonella Typhimurium that contained invA and spvC genes are pathogenic and virulent strains.</jats:p>

Identification and bioinformatic analysis of invA gene of Salmonella in free range chicken

“…Using conventional PCR approach, 9 out of 36 of microbiologically Salmonella positive isolates were detected invA-positive in free range chicken which is in agreement with the results of previous studies suggested that PCR amplification of invA gene made a reliable rapid diagnosis of salmonella spp. in the samples compared with samples as evaluated by the traditional cultural and biochemical methods (El-Sharkawy et al, 2017;Siala et al, 2017) in pig, ruminant and chicken (Scholz et al, 2001;Al-Iedani et al, 2015;Akinola et al, 2019;Mkangara et al, 2020). In the present study, the overall rate of Salmonella isolates in the free-range chicken was significantly lower than that of the commercial raised chicken (Harvey et al, 2011;Jajere et al, 2019).…”

Aplikasi Basic Local Alignment Search Tool (BLAST) NCBI Pada Penelitian Molekuler Salmonella SPP

“…BLASTn merupakan fitur pada BLAST yang dapat membandingkan suatu query sequence yang diinginkan dengan database sekuen nukleotida yang ada pada database NCBI (Hennebry et al, 2012). Penelitian yang mengaplikasikan fungsi ini antara lain dilakukan oleh Mkangara et al (2020) menggunakan BLASTn untuk mengidentifikasi informasi serovar pada isolat ayam kampung, hasil BLAST menunjukkan isolat merupakan Salmonella enterica serovar Typhimurium strain 14028 dan Salmonella enterica subspecies enterica serovar Typhimurium strain 14028. Bano et al (Hung & Weng, 2016).…”

“…Gen target yang banyak digunakan pada penelitian deteksi dan identifikasi adalah gen invasif InvA (Melati, Nurjanah, & Rahayu, 2022). Penggunaan gen ini sebagai target potensial adalah karena gen ini ditemukan spesifik pada strain Salmonella (Mkangara, Mbega, & Chacha, 2020). Selain menggunakan gen invA Penggunaan gen target lain dilakukan Nurjanah et al (2018) dengan menggunakan gen STM dan plasmid virulen Prot6E spesifik untuk Salmonella Typhimurium dan Salmonella Enteritidis dan tidak mendeteksi serovar lain (Muhsinin, Sulastri, & Supriadi, 2019).…”