Rajasekharan S K, Ray A K, Ramesh S, Kannappan Mohanvel S
Centre for Research and Development, PRIST University, Thanjavur, India.
School of Chemical Engineering, Yeungnam University, Gyeongsan, Republic of Korea.
Lett Appl Microbiol. 2018 May;66(5):378-383. doi: 10.1111/lam.12861. Epub 2018 Mar 7.
Inteins (internal proteins) are self-splicing transportable genetic elements present in conserved regions of housekeeping genes. The study highlights the importance of intein as a potential diagnostic marker for species-specific identification of Candida tropicalis, a rapidly emerging opportunistic human pathogen. Initial steps of primer validation, sequence alignment, phylogenetic tree analysis, gel electrophoresis and real-time polymerase chain reaction (PCR) assays were performed to confirm the specificity of the designed primers. The primers were selective for C. tropicalis with 100% inclusivity and showed no cross-species or cross-genera matches. The established technique is a prototype for developing multifaceted PCR assays and for point-of-care testing in near future.
Development of molecular markers for specific detection of microbial pathogens using real-time polymerase chain reaction (PCR) is an appealing and challenging technique. A real-time PCR is an emerging technology frequently used to detect the aetiologic agents. In recent times, designing species-specific primers for pathogen detection is gaining momentum. The method offers rapid, accurate and cost-effective strategy to identify the target, thus providing sufficient time to instigate appropriate chemotherapy. The study highlights the use of intein DNA sequence as molecular markers for species-specific identification of Candida tropicalis. The study also offers a prototype model for developing multifaceted PCR assays using intein DNA sequences, and provides a developmental starting point for point-of-care testing in near future.
内含肽（内部蛋白质）是存在于管家基因保守区域的自我剪接可移动遗传元件。该研究强调了内含肽作为热带假丝酵母菌物种特异性鉴定潜在诊断标志物的重要性，热带假丝酵母菌是一种迅速出现的机会性人类病原体。进行了引物验证、序列比对、系统发育树分析、凝胶电泳和实时聚合酶链反应（PCR）检测的初步步骤，以确认所设计引物的特异性。这些引物对热带假丝酵母菌具有选择性，包容性为100%，未显示跨物种或跨属匹配。所建立的技术是开发多方面PCR检测方法以及在不久的将来进行即时检测的原型。
利用实时聚合酶链反应（PCR）开发用于特异性检测微生物病原体的分子标志物是一项有吸引力且具有挑战性的技术。实时PCR是一种常用于检测病原体的新兴技术。近年来，设计用于病原体检测的物种特异性引物正成为一种趋势。该方法提供了一种快速、准确且经济高效的策略来识别目标，从而为启动适当的化疗提供足够的时间。该研究强调了内含肽DNA序列作为热带假丝酵母菌物种特异性鉴定的分子标志物的用途。该研究还提供了一个利用内含肽DNA序列开发多方面PCR检测方法的原型模型，并为不久的将来的即时检测提供了一个发展起点。
