TY - JOUR
T1 - New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay
AU - Sunarno,
AU - Khariri,
AU - Muna, Fauzul
AU - Sariadji, Kambang
AU - Rukminiati, Yuni
AU - Febriyana, Dwi
AU - Febrianti, Tati
AU - Saraswati, Ratih Dian
AU - Susanti, Ida
AU - Puspandari, Nelly
AU - Karuniawati, Anis
AU - Malik, Amarila
AU - Soebandrio, Amin
N1 - Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/5
Y1 - 2021/5
N2 - In diphtheria laboratory examinations, the PCR test can be applied to isolates and clinical specimens. This study aimed to develop a PCR assay to identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types. Seven reference isolates, four synthetic DNA samples, 36 stored isolates, and 487 clinical samples used for PCR optimization. The PCR results was confirmed by DNA sequence analysis. The results of the PCR examination of the 7 reference isolates and 36 stored isolates were similar to the results obtained using conventional methods as gold standard, both for diphtheria-causing and non-diphtheria-causing bacteria. The validation of the PCR results using DNA sequence analysis showed that there was no mispriming or misamplification. The multiplex PCR assay developed in this study could correctly identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types not yet covered by established PCR methods.
AB - In diphtheria laboratory examinations, the PCR test can be applied to isolates and clinical specimens. This study aimed to develop a PCR assay to identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types. Seven reference isolates, four synthetic DNA samples, 36 stored isolates, and 487 clinical samples used for PCR optimization. The PCR results was confirmed by DNA sequence analysis. The results of the PCR examination of the 7 reference isolates and 36 stored isolates were similar to the results obtained using conventional methods as gold standard, both for diphtheria-causing and non-diphtheria-causing bacteria. The validation of the PCR results using DNA sequence analysis showed that there was no mispriming or misamplification. The multiplex PCR assay developed in this study could correctly identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types not yet covered by established PCR methods.
KW - Diphtheria
KW - dtxR gene
KW - PCR
KW - Potentially toxigenic Corynebacterium
UR - http://www.scopus.com/inward/record.url?scp=85103000221&partnerID=8YFLogxK
U2 - 10.1016/j.mimet.2021.106198
DO - 10.1016/j.mimet.2021.106198
M3 - Article
C2 - 33713727
AN - SCOPUS:85103000221
VL - 184
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
SN - 0167-7012
M1 - 106198
ER -