Establishment of a recombined mannose-binding lectin protein-magnetic beads-enriched binding recombinant enzyme-assisted polymerase chain reaction assay for Candida in blood samples.
10.3760/cma.j.cn112338-20230215-00079
- VernacularTitle:血液中念珠菌重组人甘露聚糖结合凝集素蛋白磁珠富集联合重组酶辅助聚合酶链式反应检测方法的建立
- Author:
Meng Yi ZHANG
1
;
Xiao Ping CHEN
2
;
Xiu Li SUN
3
;
Xue Jun MA
4
;
Xin Xin SHEN
4
;
Yan Yan GUO
5
Author Information
1. College of Clinical Medicine, North China University of Science and Technology, Tangshan 063210, China National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China Clinical Laboratory, Tangshan Gongren Hospital, Tangshan 063000, China.
2. National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
3. College of Clinical Medicine, North China University of Science and Technology, Tangshan 063210, China National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
4. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
5. Clinical Laboratory, Tangshan Gongren Hospital, Tangshan 063000, China.
- Publication Type:Journal Article
- MeSH:
Humans;
Lectins;
Candida;
Candidemia;
Reproducibility of Results;
Polymerase Chain Reaction;
Nucleic Acids;
Magnetic Phenomena
- From:
Chinese Journal of Epidemiology
2023;44(5):823-827
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish a nested recombinant enzyme-assisted polymerase chain reaction (RAP) technique combined with recombined mannose-binding lectin protein (M1 protein)-magnetic beads enrichment for the detection of Candida albicans (C. albicans) and Candida tropicalis (C. tropicalis) in blood samples for the early diagnosis of candidemia albicans and candidiemia tropicalis. Methods: The primer probes for highly conserved regions of the internal transcribed spacerregions of C. albicans and C. tropicalis were deigned to establish RAP assays for the detections of C. albicans and C. tropicalis; The sensitivity and reproducibility of nucleic acid tests with gradient dilutions of standard strains and specificity of nucleic acid tests with common clinical pathogens causing bloodstream infection were condcuted. M1 protein-magnetic bead enriched plasma C. albicans and C. tropicalis were used for RAP and PCR in with simulated samples and the results were compared. Results: The sensitivity of the established dual RAP assay was 2.4-2.8 copies/reaction, with higher reproducibility and specificity. M1 protein-magnetic bead enrichment of pathogen combined with the dual RAP assay could complete the detections of C. albicans and C. tropicalis in plasma within 4 hours. Fie the pathogen samples at concentration <10 CFU/ml, the number of the samples tested by RAP was higher than that tested by PCR after enrichment. Conclusion: In this study, a dual RAP assay for the detections of C. albicans and C. tropicalis in blood sample was developed, which has the advantages of accuracy, rapidity, and less contaminants and has great potential for rapid detection of Candidemia.
