Assay for detection of toxigenic Clostridioides difficile with combined microfluidic chip and immunochromatography technology
10.3969/j.issn.1002-2694.2025.00.031
- VernacularTitle:基于微流控芯片PCR结合免疫层析技术检测产毒型艰难梭菌方法的建立与性能评价
- Author:
Hong-rui CHENG
1
;
Xiao-jun SONG
;
Yu CHEN
;
Meng ZHANG
;
Meng-ting CAI
;
Kun ZHU
;
Yu-lei TAI
;
Shi-bo YING
;
Da-zhi JIN
Author Information
1. 杭州医学院,检验医学院、生物工程学院,杭州 310053
- Publication Type:Journal Article
- Keywords:
Clostridioides difficile;
microfluidic chip;
immunochromatography;
detection
- From:
Chinese Journal of Zoonoses
2025;41(2):142-149
- CountryChina
- Language:Chinese
-
Abstract:
An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.
