Ranjbar Reza, Mortazavi Seyyed Mojtaba, Mehrabi Tavana Ali, Sarshar Meysam, Najafi Ali, Soruri Zanjani Rahim
Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Dept. of Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Iran J Public Health. 2017 Jan;46(1):103-111.
serovar Typhi, as causative agent of typhoid fever, is one of the most important endemic pathogens. Non-typhoidal serovars, including Typhimurium, Infantis, and Enteritidis are amongst the most prevalent serotypes worldwide and in developing areas such as Iran. The aim of this study was to apply a uniplex PCR for rapid detection of spp., and a multiplex PCR for the simultaneous detection of the four most common serovars in Iran.
Current research was done in 2010 at Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. For detection of spp a pair of primers was used to replicate a chromosomal sequence. Four other sets of primers were also designed to amplify the target genes of four species including , and three non-typhoidal spp (). The assay specificity was investigated by testing 15 different serovars and 8 other additional non- species.
The genus-specific PCR yielded the expected DNA band of 404 bp in all spp., strains tested. The uniplex and multiplex PCR assays produced also the expected fragments of 489 bp, 304 bp, 224 bp, and 104 bp for serovars Typhi, Enteritidis, Typhimurium, and Infantis, respectively. Each species-specific primer pair set did not show any cross-reactivity when tested on other serovars or other non- but related- strains.
Both uniplex and multiplex PCR protocols had a good specificity. They can provide an important tool for the rapid and simultaneous detection and differentiation of the four most prevalent serovars in Iran.
伤寒血清型作为伤寒热的病原体，是最重要的地方性病原菌之一。非伤寒血清型，包括鼠伤寒血清型、婴儿血清型和肠炎血清型，是全球以及伊朗等发展中地区最普遍的血清型。本研究的目的是应用单重聚合酶链反应（PCR）快速检测沙门氏菌属，并应用多重PCR同时检测伊朗四种最常见的沙门氏菌血清型。
当前研究于2010年在伊朗德黑兰巴基耶塔拉医科大学分子生物学研究中心进行。为检测沙门氏菌属，使用一对引物复制一个染色体序列。还设计了另外四组引物来扩增四种沙门氏菌的靶基因，包括伤寒沙门氏菌，以及三种非伤寒沙门氏菌血清型（鼠伤寒沙门氏菌、婴儿沙门氏菌和肠炎沙门氏菌）。通过检测15种不同的沙门氏菌血清型和8种其他非沙门氏菌属菌株来研究该检测方法的特异性。
在所有检测的沙门氏菌属菌株中，属特异性PCR产生了预期的404 bp DNA条带。单重和多重PCR检测还分别产生了伤寒血清型、肠炎血清型、鼠伤寒血清型和婴儿血清型预期的489 bp、304 bp、224 bp和104 bp片段。当在其他沙门氏菌血清型或其他非沙门氏菌但相关的沙门氏菌菌株上进行测试时，每种物种特异性引物对均未显示任何交叉反应。
单重和多重PCR方案均具有良好的特异性。它们可为快速、同时检测和区分伊朗四种最普遍的沙门氏菌血清型提供重要工具。
