PDF(Pubmed)
Abstract：
The study aimed to develop a quantitative colorimetric loop-mediated isothermal amplification technique using the phenol red indicator (QLAMP-PhR) for detecting Fusobacterium nucleatum (Fn) levels in colorectal cancer (CRC) patients and healthy individuals. QLAMP-PhR assays were conducted on 251 stool samples specific for the Fn FadA gene. Six primers were synthesized and utilized with master mix reagents, and a phenol red indicator was employed to enhance the QLAMP-PhR technique. A standard quantitative analysis curve was generated using a logarithmic function (absorbance vs. concentration) by serially diluting the copy number of genomic DNA templates (Fn ATCC25586). The CRC group exhibited a significantly higher abundance of Fn compared to the healthy control group (P < 0.001). These findings suggest that the QLAMP-PhR technique effectively identifies Fn specifically by its gene for the key virulence factor FadA. Additionally, ideas for developing a real-time QLAMP-PhR test were presented. Compared to the traditional polymerase chain reaction (PCR) technique, QLAMP-PhR offers several advantages including rapidity, simplicity, specificity, sensitivity, and cost-effectiveness method that can quantitatively screen for Fn presence in normal populations. The QLAMP-PhR method represents a sensitive and specific amplification assay for the rapid detection of the Fn pathogen. To the best of our knowledge, this study is the first to report the application of QLAMP-PhR for detecting FadA in Fn.
摘要：
该研究旨在开发一种使用酚红指示剂（QLAMP-PhR）的定量比色环介导等温扩增技术，用于检测结直肠癌（CRC）患者和健康个体中核梭杆菌（Fn）的水平。对FnFadA基因特异性的251个粪便样品进行QLAMP-PhR测定。合成了六种引物，并与主混合试剂一起使用，使用酚红指示剂来增强QLAMP-PhR技术。使用对数函数（吸光度与浓度）通过连续稀释基因组DNA模板的拷贝数（FnATCC25586）。与健康对照组相比,CRC组表现出显著更高的Fn丰度(P<0.001)。这些发现表明，QLAMP-PhR技术通过其关键毒力因子FadA的基因有效地鉴定了Fn。此外,提出了开发实时QLAMP-PhR测试的思路。与传统的聚合酶链反应(PCR)技术相比,QLAMP-PhR提供了几个优点，包括快速性，简单，特异性，灵敏度,和成本效益方法，可以定量筛选正常人群中Fn的存在。QLAMP-PhR方法代表了快速检测Fn病原体的灵敏和特异性扩增测定。据我们所知,本研究首次报道了QLAMP-PhR在Fn中检测FadA的应用。
