Yang Yuanyuan, Rajendran Vinod, Jayaraman Vasanth, Wang Tianhao, Bei Kang, Krishna Karthik, Rajasekaran Karenah, Rajasekaran John J, Krishnamurthy Hari
Vibrant America LLC, San Carlos, CA USA.
Vibrant Sciences LLC, San Carlos, CA USA.
Gut Pathog. 2019 Oct 18;11:51. doi: 10.1186/s13099-019-0329-2. eCollection 2019.
Rapid detection of a wide range of etiologic agents is essential for appropriate treatment and control of gastrointestinal (GI) infections. A variety of microbial species including bacteria, viruses, parasites, and fungi have been recognized as diarrheagenic enteric pathogens. However, multiplex testing of various targets in a single reaction needs further improvement because of its limitation in species and throughput.
This study aims at developing and evaluating a DNA microarray-based qualitative multiplexed polymerase chain reaction (PCR) assay, Vibrant GI pathogen panel (GPP), for simultaneous detection of 27 enteric GI pathogenic targets (16 bacteria, 5 viruses, 4 parasites, and 2 fungi) directly from stool specimens. Limits of detection ranged from 10 to 10 cells/mL for bacteria, 10 to 10 cells/mL for parasites, 10 to 10 RNA copies/mL for viruses, and 10 to 10 cells/mL for fungi. Performance characteristics were determined using 27 Quantitative Genomic DNAs, 212 spiked stool specimens, 1067 clinical and archived stool specimens. Overall sensitivity was 95.9% (95% CI 92.4-98.1) and specificity was 100% (95% CI 99.9-100). Polymicrobial detections contained either two or three organisms was 20.2% (35/173) of positive clinical specimens and 3.3% (35/1055) of all clinical specimens.
The Vibrant GPP is a comprehensive, high-throughput, and rapid DNA microarray to provide etiologic diagnosis of GI infections in the laboratory setting.
快速检测多种病原体对于胃肠道（GI）感染的恰当治疗和控制至关重要。包括细菌、病毒、寄生虫和真菌在内的多种微生物已被确认为致泻性肠道病原体。然而，由于其在物种和通量方面的局限性，单一反应中对各种靶标的多重检测仍需进一步改进。
本研究旨在开发和评估一种基于DNA微阵列的定性多重聚合酶链反应（PCR）检测方法，即活力胃肠道病原体检测板（GPP），用于直接从粪便标本中同时检测27种肠道GI致病靶标（16种细菌、5种病毒、4种寄生虫和2种真菌）。细菌的检测限为10至10细胞/毫升，寄生虫为10至10细胞/毫升，病毒为10至10 RNA拷贝/毫升，真菌为10至10细胞/毫升。使用27种定量基因组DNA、212份加标粪便标本、1067份临床和存档粪便标本确定了性能特征。总体敏感性为95.9%（95%CI 92.4 - 98.1），特异性为100%（95%CI 99.9 - 100）。包含两种或三种微生物的多微生物检测在阳性临床标本中占20.2%（35/173），在所有临床标本中占3.3%（35/1055）。
活力GPP是一种全面、高通量且快速的DNA微阵列，可在实验室环境中为GI感染提供病因诊断。
