Simultaneous detection and characterization of toxigenic Clostridium difficile directly from clinical stool specimens.
10.1007/s11684-017-0560-5
- Author:
Hanjiang LAI
1
;
Chen HUANG
2
;
Jian CAI
3
;
Julian YE
2
;
Jun SHE
1
;
Yi ZHENG
4
;
Liqian WANG
5
;
Yelin WEI
1
;
Weijia FANG
4
;
Xianjun WANG
5
;
Yi-Wei TANG
6
;
Yun LUO
7
;
Dazhi JIN
8
Author Information
1. The First People's Hospital of Xiaoshan District, Hangzhou, 311021, China.
2. Department of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China.
3. Department of Disease Control and Prevention, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China.
4. Biotherapy Center for Medical Oncology, the First Affiliated Hospital, Zhejiang University, Hangzhou, 310003, China.
5. Department of Laboratory Medicine, Hangzhou First People's Hospital, Hangzhou, 310006, China.
6. Department of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA.
7. Department of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China. yluo@cdc.zj.cn.
8. Department of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, 310051, China. dzjin@cdc.zj.cn.
- Publication Type:Journal Article
- Keywords:
Clostridium difficile;
capillary electrophoresis;
characterization;
detection;
multiplex PCR
- MeSH:
Clostridium Infections;
diagnosis;
Clostridium difficile;
genetics;
Electrophoresis, Capillary;
Feces;
microbiology;
Genes, Bacterial;
Humans;
Polymerase Chain Reaction;
Ribotyping;
Sensitivity and Specificity
- From:
Frontiers of Medicine
2018;12(2):196-205
- CountryChina
- Language:English
-
Abstract:
We employed a multiplex polymerase chain reaction (PCR) coupled with capillary electrophoresis (mPCR-CE) targeting six Clostridium difficile genes, including tpi, tcdA, tcdB, cdtA, cdtB, and a deletion in tcdC for simultaneous detection and characterization of toxigenic C. difficile directly from fecal specimens. The mPCR-CE had a limit of detection of 10 colony-forming units per reaction with no cross-reactions with other related bacterial genes. Clinical validation was performed on 354 consecutively collected stool specimens from patients with suspected C. difficile infection and 45 isolates. The results were compared with a reference standard combined with BD MAX Cdiff, real-time cell analysis assay (RTCA), and mPCR-CE. The toxigenic C. difficile species were detected in 36 isolates and 45 stool specimens by the mPCR-CE, which provided a positive rate of 20.3% (81/399). The mPCR-CE had a specificity of 97.2% and a sensitivity of 96.0%, which was higher than RTCA (x = 5.67, P = 0.017) but lower than BD MAX Cdiff (P = 0.245). Among the 45 strains, 44 (97.8%) were determined as nonribotype 027 by the mPCR-CE, which was fully agreed with PCR ribotyping. Even though ribotypes 017 (n = 8, 17.8%), 001 (n = 6, 13.3%), and 012 (n = 7, 15.6%) were predominant in this region, ribotype 027 was an important genotype monitored routinely. The mPCR-CE provided an alternative diagnosis tool for the simultaneous detection of toxigenic C. difficile in stool and potentially differentiated between RT027 and non-RT027.
