Deakin University, Geelong, Australia.
Barwon Health, Geelong, Australia.
J Dev Orig Health Dis. 2020 Jun;11(3):228-234. doi: 10.1017/S2040174419000849. Epub 2019 Dec 17.
Gut bacteria from the genus Prevotella are found in high abundance in faeces of non-industrialised communities but low abundance in industrialised, Westernised communities. Prevotella copri is one of the principal Prevotella species within the human gut. As it has been associated with developmental health and disease states, we sought to (i) develop a real-time polymerase chain reaction (PCR) to rapidly determine P. copri abundance and (ii) investigate its abundance in a large group of Australian pregnant mothers.The Barwon Infant Study is a pre-birth cohort study (n = 1074). Faecal samples were collected from mothers at 36 weeks gestation. Primers with a probe specific to the V3 region of P. copri 16S rRNA gene were designed and optimised for real-time PCR. Universal 16S rRNA gene primers amplified pan-bacterial DNA in parallel. Relative abundance of P. copri was calculated using a 2-ΔCt method.Relative abundance of P. copri by PCR was observed in 165/605 (27.3%) women. The distribution was distinctly bimodal, defining women with substantial (n = 115/165, 69.7%) versus very low P. copri expression (n = 50/165, 30.3%). In addition, abundance of P. copri by PCR correlated with 16S rRNA gene MiSeq sequencing data (r2 = 0.67, P < 0.0001, n = 61).We have developed a rapid and cost-effective technique for identifying the relative abundance of P. copri using real-time PCR. The expression of P. copri was evident in only a quarter of the mothers, and either at substantial or very low levels. PCR detection of P. copri may facilitate assessment of this species in large, longitudinal studies across multiple populations and in various clinical settings.
瘤胃丙酸杆菌属的肠道细菌在非工业化社区的粪便中大量存在,但在工业化、西化的社区中含量较低。普雷沃氏菌是人类肠道中主要的普雷沃氏菌属之一。由于它与发育健康和疾病状态有关,我们试图(i)开发一种实时聚合酶链反应(PCR)来快速确定 P. copri 的丰度,(ii)在一大群澳大利亚孕妇中研究其丰度。巴旺婴儿研究是一项产前队列研究(n = 1074)。在 36 周妊娠时从母亲那里收集粪便样本。设计并优化了针对 P. copri 16S rRNA 基因 V3 区的引物和探针,用于实时 PCR。通用 16S rRNA 基因引物平行扩增泛细菌 DNA。使用 2-ΔCt 方法计算 P. copri 的相对丰度。PCR 检测到 165/605(27.3%)名女性中有 P. copri。分布明显呈双峰型,定义为大量(n = 115/165,69.7%)与极低 P. copri 表达(n = 50/165,30.3%)的女性。此外,PCR 检测到的 P. copri 丰度与 16S rRNA 基因 MiSeq 测序数据相关(r2 = 0.67,P < 0.0001,n = 61)。我们开发了一种使用实时 PCR 快速、经济高效地识别 P. copri 相对丰度的技术。只有四分之一的母亲表达了 P. copri,而且表达水平要么很高,要么很低。PCR 检测 P. copri 可能有助于在多个人群和各种临床环境中进行的大型纵向研究中评估该物种。