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Comparison of Conventional PCR Multiplex PCR and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

机译:常规PCR多重PCR和环介导的等温扩增测定法用于快速检测Arcobacter种的比较

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摘要

This study aimed to develop a loop-mediated isothermal amplification (LAMP) method for the rapid detection of Arcobacter species. Specific primers targeting the 23S ribosomal RNA gene were used to detect Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The specificity of the LAMP primer set was assessed using DNA samples from a panel of Arcobacter and Campylobacter species, and the sensitivity was determined using serial dilutions of Arcobacter species cultures. LAMP showed a 10- to 1,000-fold-higher sensitivity than multiplex PCR, with a detection limit of 2 to 20 CFU per reaction in vitro. Whereas multiplex PCR showed cross-reactivity with Campylobacter species, the LAMP method developed in this study was more sensitive and reliable than conventional PCR or multiplex PCR for the detection of Arcobacter species.
机译:这项研究旨在开发一种环介导的等温扩增(LAMP)方法,用于快速检测Arcobacter菌种。靶向23S核糖体RNA基因的特异引物用于检测Butzleri弓形杆菌,cryalerophilus弓形杆菌和skirrowii杆菌。 LAMP引物组的特异性是使用来自一组杆状杆菌和弯曲杆菌属物种的DNA样品评估的,敏感性是使用杆状杆菌属培养物的系列稀释液确定的。 LAMP显示的灵敏度比多重PCR高10到1,000倍,体外每个反应的检测极限为2到20 CFU。尽管多重PCR显示与弯曲杆菌属物种具有交叉反应性,但本研究中开发的LAMP方法比常规PCR或多重PCR更灵敏和可靠。

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