New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

Sunarno; Khariri; Fauzul Muna; Kambang Sariadji; Yuni Rukminiati; Dwi Febriyana; Tati Febrianti; Ratih Dian Saraswati; Ida Susanti; Nelly Puspandari; Anis Karuniawati; Amarila Malik; Amin Soebandrio

doi:10.1016/j.mimet.2021.106198

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

Sunarno, Khariri, Fauzul Muna, Kambang Sariadji, Yuni Rukminiati, Dwi Febriyana, Tati Febrianti, Ratih Dian Saraswati, Ida Susanti, Nelly Puspandari, Anis Karuniawati, Amarila Malik, Amin Soebandrio

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1 Citation (Scopus)

Abstract

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.

Original language	English
Article number	106198
Journal	Journal of Microbiological Methods
Volume	184
DOIs	https://doi.org/10.1016/j.mimet.2021.106198
Publication status	Published - May 2021

Keywords

Diphtheria
dtxR gene
PCR
Potentially toxigenic Corynebacterium

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10.1016/j.mimet.2021.106198

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Sunarno, Khariri, Muna, F., Sariadji, K., Rukminiati, Y., Febriyana, D., Febrianti, T., Saraswati, R. D., Susanti, I., Puspandari, N., Karuniawati, A., Malik, A., & Soebandrio, A. (2021). New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay. Journal of Microbiological Methods, 184, [106198]. https://doi.org/10.1016/j.mimet.2021.106198

Sunarno ; Khariri ; Muna, Fauzul ; Sariadji, Kambang ; Rukminiati, Yuni ; Febriyana, Dwi ; Febrianti, Tati ; Saraswati, Ratih Dian ; Susanti, Ida ; Puspandari, Nelly ; Karuniawati, Anis ; Malik, Amarila ; Soebandrio, Amin. / New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay. In: Journal of Microbiological Methods. 2021 ; Vol. 184.

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title = "New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay",

abstract = "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.",

keywords = "Diphtheria, dtxR gene, PCR, Potentially toxigenic Corynebacterium",

author = "Sunarno and Khariri and Fauzul Muna and Kambang Sariadji and Yuni Rukminiati and Dwi Febriyana and Tati Febrianti and Saraswati, {Ratih Dian} and Ida Susanti and Nelly Puspandari and Anis Karuniawati and Amarila Malik and Amin Soebandrio",

note = "Publisher Copyright: {\textcopyright} 2021 Elsevier B.V.",

year = "2021",

month = may,

doi = "10.1016/j.mimet.2021.106198",

language = "English",

volume = "184",

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Sunarno, Khariri, Muna, F, Sariadji, K, Rukminiati, Y, Febriyana, D, Febrianti, T, Saraswati, RD, Susanti, I, Puspandari, N, Karuniawati, A , Malik, A & Soebandrio, A 2021, 'New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay', Journal of Microbiological Methods, vol. 184, 106198. https://doi.org/10.1016/j.mimet.2021.106198

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay. / Sunarno; Khariri; Muna, Fauzul; Sariadji, Kambang; Rukminiati, Yuni; Febriyana, Dwi; Febrianti, Tati; Saraswati, Ratih Dian; Susanti, Ida; Puspandari, Nelly; Karuniawati, Anis ; Malik, Amarila ; Soebandrio, Amin.

In: Journal of Microbiological Methods, Vol. 184, 106198, 05.2021.

Research output: Contribution to journal › Article › peer-review

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

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

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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 -

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

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