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Detection of virulence genes of Clostridium difficile by multiplex PCR
Author(s) -
ANTIKAINEN JENNI,
PASANEN TANJA,
MERO SOINTU,
TARKKA EVELIINA,
KIRVESKARI JUHA,
KOTILA SAARA,
MENTULA SILJA,
KÖNÖNEN EIJA,
VIROLAINENJULKUNEN ANNIRIITTA,
VAARA MARTTI,
TISSARI PÄIVI
Publication year - 2009
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1111/j.1600-0463.2009.02509.x
Subject(s) - clostridium difficile , multiplex polymerase chain reaction , virulence , microbiology and biotechnology , biology , clostridium difficile toxin a , pulsed field gel electrophoresis , ribotyping , polymerase chain reaction , pore forming toxin , clostridium difficile toxin b , multiplex , variants of pcr , clostridiaceae , virology , typing , toxin , gene , genotype , genetics , microbial toxins , antibiotics
Antikainen J, Pasanen T, Mero S, Tarkka E, Kirveskari J, Kotila S, Mentula S, Könönen E, Virolainen‐Julkunen A‐R, Vaara M, Tissari P. Detection of virulence genes of Clostridium difficile by multiplex PCR. APMIS 2009; 117: 607–13. Clostridium difficile strains belonging to the PCR ribotype 027, pulse‐field gel electrophoresis (PFGE) type NAP1, toxinotype III and restriction endonuclease analysis group BI harbouring mutations in the tcdC gene and possessing binary toxin components A and B have been described to cause epidemics with increased morbidity and mortality. In the present study we developed a conventional multiplex PCR designed to detect selected virulence associated markers of the hypervirulent C. difficile PCR ribotype 027. The multiplex PCR assay detected the major toxins A and B, binary toxin components A and B as well as a possible deletion in the tcdC gene: a characteristic pattern of amplification products for the PCR ribotype 027 strains was detected. This rather simple method was specific for the screening of this hypervirulent C. difficile strain. The correlation between the multiplex PCR and PCR ribotyping methods was excellent. The sensitivity and specificity were 100% in our epidemiological situation. In conclusion, this multiplex PCR was found useful in the preliminary screening for the hypervirulent C. difficile PCR ribotype 027.