Glover Quarshie, Jiang Xiao, Onderak Alexis Marie, Mapes Abigail, Hollnagel Fauzia, Buckley Joseph, Kim Chang Hee, Siraj Dawd
Department of Medicine, University of Wisconsin Hospitals and Clinics, Madison, Wisconsin, United States of America.
PLoS One. 2025 Mar 12;20(3):e0319145. doi: 10.1371/journal.pone.0319145. eCollection 2025.
Infectious diarrheal diseases are one of the leading causes of worldwide morbidity and mortality. The incidence of diarrhea is higher in Low-Middle-Income Countries (LMIC), where more than 90% of deaths from diarrheal diseases occur. Diagnostic tests for infectious diarrhea are not readily available in Low-Middle-Income Countries. Our study evaluates a novel, cost-effective, easy-to-use DNA stool testing kit for infectious diarrhea that can easily be rolled out in low-resource settings and has comparable performance to current testing modalities in the USA in terms of diagnostic utility.
435 stool samples were tested using the novel stool testing Kit (Go-GutDx®). The stool samples were in groups of 8 and were tested using floating microspheres DNA extraction followed by Recombinase Polymerase Amplification (RPA) and lateral flow assay detection. Pathogens tested include Clostridium difficile, Campylobacter jejuni, Salmonella enterica Typhimurium, Shigella spp., STEC (stx1, stx2), Vibrio spp., and Yersinia enterocolitica. The same samples were tested using BioFire GI Panel (gold standard), and the results were compared. Descriptive analysis was summarized as raw counts and frequencies and compared using Fisher's exact test. We conducted specificity and sensitivity analysis of the two diagnostic tests using the diagnostic testing package in STATA. Graphical illustrations were conducted using two-way line graphs with custom margins and axis. All analyses with p-values ≤ 0.05 were significant. All analyses were conducted using STATA version 17.
Results showed a sensitivity of 56.1% for C. difficile, and 58.6% for Campylobacter jejuni. 46.1% for STEC (stx1, stx2), 83.3% for Salmonella, 0% for Yersinia enterocolitica, 66.7% for Vibrio spp and 65.0% for Shigella spp. Specificities were > 97% for all pathogens. The positive predictive value was highest for Campylobacter jejuni 94.4%, followed by C. difficile 93.5%, Salmonella 86.2%, Shigella 76.5%, STEC (stx1, stx2) 50%, Vibrio spp 40%, and Yersinia 0%. The negative predictive value was 75.8% for C. difficile, with all other pathogens being above 98.3%. Table 3. The concordance between BioFire and Go-GutDx® for C. difficile testing was 80%. All other organisms showed a concordance of greater than 97% Table 2.
Our study confirms that Go-GutDx® is a novel diagnostic tool for diarrheal pathogens with statistically comparable sensitivity, specificity, positive and negative predictive values with the current gold standard testing kit, BioFire. In addition, the simplicity of the technology and lower price both to healthcare systems and to patients makes Go-GutDx® an ideal diagnostic test to be implemented in low- and middle-income countries.
感染性腹泻病是全球发病和死亡的主要原因之一。腹泻发病率在低收入和中等收入国家（LMIC）较高，超过90%的腹泻病死亡发生在这些国家。低收入和中等收入国家难以获得感染性腹泻的诊断检测方法。我们的研究评估了一种新型、经济高效、易于使用的用于感染性腹泻的粪便DNA检测试剂盒，该试剂盒可在资源匮乏地区轻松推广，并且在诊断效用方面与美国目前的检测方法具有相当性能。
使用新型粪便检测试剂盒（Go-GutDx®）对435份粪便样本进行检测。粪便样本以8份为一组，采用浮动微球DNA提取法进行检测，随后进行重组酶聚合酶扩增（RPA）和侧向流动分析检测。检测的病原体包括艰难梭菌、空肠弯曲菌、鼠伤寒沙门氏菌、志贺氏菌属、肠出血性大肠杆菌（stx1、stx2）、弧菌属和小肠结肠炎耶尔森菌。使用BioFire GI Panel（金标准）对相同样本进行检测，并比较结果。描述性分析总结为原始计数和频率，并使用Fisher精确检验进行比较。我们使用STATA中的诊断测试包对两种诊断测试进行特异性和敏感性分析。使用带有自定义边距和轴的双向线图进行图形说明。所有p值≤0.05的分析均具有显著性。所有分析均使用STATA 17版本进行。
结果显示，艰难梭菌的敏感性为56.1%，空肠弯曲菌为58.6%。肠出血性大肠杆菌（stx1、stx2）为46.1%，沙门氏菌为83.3%，小肠结肠炎耶尔森菌为0%，弧菌属为66.7%，志贺氏菌属为65.0%。所有病原体的特异性均>97%。空肠弯曲菌的阳性预测值最高，为94.4%，其次是艰难梭菌，为93.5%，沙门氏菌为86.2%，志贺氏菌为76.5%，肠出血性大肠杆菌（stx1、stx2）为50%，弧菌属为40%，耶尔森菌为0%。艰难梭菌的阴性预测值为75.8%，其他所有病原体均高于98.3%。表3。BioFire和Go-GutDx®在艰难梭菌检测方面的一致性为80%。所有其他生物体的一致性均大于97% 表2。
我们的研究证实，Go-GutDx®是一种用于腹泻病原体的新型诊断工具，在统计学上，其敏感性、特异性、阳性和阴性预测值与当前的金标准检测试剂盒BioFire相当。此外，该技术的简单性以及对医疗系统和患者而言较低的价格，使得Go-GutDx®成为在低收入和中等收入国家实施的理想诊断测试。
