PCR coupled with mass-spectrometry for detection of Clostridium difficile virulence markers during the emergence of ribotype 027 in Bucharest area

被引:1
作者
Florea, Dragos [1 ,3 ]
Huhulescu, Steliana [2 ]
Indra, Alexander [2 ]
Badicut, Ioana [1 ]
Rafila, Alexandru [1 ,3 ]
Otelea, Dan [1 ]
Popescu, Gabriel Adrian [1 ,3 ]
机构
[1] Natl Inst Infect Dis Prof Dr Matei Bals, Bucharest, Romania
[2] Austrian Agcy Hlth & Food Safety AGES Vienna, Vienna, Austria
[3] Univ Med & Pharm Carol Davila, Bucharest, Romania
来源
REVISTA ROMANA DE MEDICINA DE LABORATOR | 2015年 / 23卷 / 04期
关键词
Clostridium difficile; ribotype; 027; binary toxin; deletion in tcdC gene; electrospray ionization mass spectrometry; FRAGMENT LENGTH POLYMORPHISM; POLYMERASE-CHAIN-REACTION; TOXIN; IDENTIFICATION; STRAIN; SURVEILLANCE; INFECTION; DISEASE; TCDC;
D O I
10.1515/rrlm-2015-0044
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
In recent years Clostridium difficile infection (CDI) has represented a serious public health issue, mainly due to the global spread of the hypervirulent strain NAP1/027/BI. The purpose of the present study was to evaluate the utility of a PCR coupled with electrospray ionization mass spectrometry (ESI-MS) commercial assay for the detection of C. difficile virulence markers. Non-duplicative C. difficile isolates from patients with CDI diagnosed in a tertiary level hospital from Bucharest were tested for toxin A, toxin B, binary toxin genes and deletion in tcdC gene using PCR/capillary gel electrophoresis and PCR/ESI-MS. The study analysed 45 non-duplicative isolates, 33 strains (73.3%) belonging to ribotype 027. The concordance between PCR/capillary gel electrophoresis and PCR/ESI-MS was 100% for toxin A gene, 97.8% for toxin B gene, 91.1% for binary toxin subunit A gene and 95.6% for binary toxin subunit B gene. The general concordance for the complete panel of markers was 88.9% but was 100% for ribotype 027 isolates. PCR/ESI-MS might be a valid method for the detection of C. difficile virulence markers, including binary toxin.
引用
收藏
页码:449 / 455
页数:7
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