DNA Sequence Analysis of dtxR Gene (Partial) of Corynebacterium diphtheriae Causing Diphtheria in Jawa and Kalimantan Islands, Indonesia

Sunarno, Sunarno; Mulyastuti, Yuanita; Puspandari, Nelly; Sariadji, Kambang

doi:10.18585/inabj.v9i2.268

Cited by 6 publications

(2 citation statements)

References 24 publications

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“…Gen dtxR dimiliki oleh semua strain C.diphtheriae, baik toksigenik maupun non toksigenik sehingga gen ini dapat digunakan sebagai penanda C. diphtheria (Sunarno et al, 2017). Sekuen fasta gen yang terdaftar dalam NCBI pada gambar 1 dilakukan analisa desain primer menggunakan Primer3Plus lalu dilanjutkan dengan studi In silico.…”

Section: Pembahasanunclassified

Desain Primer untuk Deteksi Gen Diphtheria Toxin Repressor (dtxR) sebagai Biomarker Bakteri Corynebacterium diphtheriae Menggunakan In Silico PCR

Nastiti,

Kuncara

2023

JLM

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Corynebacterium diphtheriae is the bacteria that causes diphtheria. The virulence factor of C. diphtheriae comes from the bacteria's ability to produce bacterial toxins. Toxin production is regulated by a set of genes called tox/dtx genes and is regulated by the dtxR gene. The aim of this study was to design primers used to evaluate the dtxR gene using bacterial DNA sequences. This research is experimental research with a literature study approach using the In silico Polymerase Chain Reaction (PCR), NCBI (National Center for Biotechnology Information), Primer3Plus, and Oligo Calculator applications. The sample obtained from genbank NCBI was C. diphtheriae dtxR gene M80337.1. In silico PCR examination was carried out using newly designed primers from Primer3Plus with 50 genomic DNA of Corynebacterium spp. taken from the In silico PCR database. The dtxR primer pair: '5-ACAGTTAGCCAAACCGTTGC-3' and 5'-TGCGTTCAACTTCGTCACTC-3' can produce a single DNA amplicon measuring 226 bp specifically for C. diphtheria types and no amplicon bands were generated from other Corynebacterium genomes. Based on the study results, this pair of specific primers can be used for in vitro PCR testing and can be used to develop rapid detection of diphtheria.

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Section: Pembahasanunclassified

Desain Primer untuk Deteksi Gen Diphtheria Toxin Repressor (dtxR) sebagai Biomarker Bakteri Corynebacterium diphtheriae Menggunakan In Silico PCR

Nastiti,

Kuncara

2023

JLM

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“…DNA was extracted using the commercial QIAamp DNA Mini Kit (Qiagen) (17). The DNA in 50 uL molecular water was stored at -20°C for the DNA template in the PCR examination.…”

Section: Multiplex Real-time Pcrmentioning

confidence: 99%

Development and Application of Dtxr and Tox Genes Targeting Real-time PCR to Identify Corynebacterium diphtheriae, C. ulcerans, and C. pseudotuberculosis Simultaneously

Sunarno¹,

Hartoyo²,

Amalia³

et al. 2022

Jundishapur J Microbiol

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Background: Corynebacterium diphtheriae, C. ulcerans, and C. pseudotuberculosis are known as diphtheria-causing bacteria. Although diphtheria therapy is administered based on the clinical manifestations, some cases are mild and atypical. The immediate and accurate identification of diphtheria-causing bacteria is of paramount importance to prevent the spread of the disease and provide case management as early as possible. Unfortunately, conventional methods as the gold standard are time-consuming. Objectives: This study aimed to develop and implement a multiplex real-time PCR with the dtxR and tox genes as the target to identify three species of diphtheria-causing bacteria and screen their toxigenicity quickly and accurately. Methods: The research sample encompassed seven reference strains, one synthetic DNA, 30 archived isolates, and 924 clinical specimens isolated from 311 diphtheria cases and 613 close contacts. The conventional methods as the gold standard and the established PCR assay were used to verify the results of multiplex real-time PCR developed in this study. Results: The multiplex real-time PCR could identify seven reference strains, one synthetic DNA, and 30 archived isolates as accurately as the conventional methods and the established PCR. Similar to established PCR, the multiplex real-time PCR identified diphtheria-causing bacteria in 120 (38.6%) out of 311 and 12 (2%) out of 613 clinical specimens from diphtheria cases and close contacts, respectively. Meanwhile, the conventional methods identified diphtheria-causing bacteria in 79 (25.4%) out of 311 and three (0.5%) out of 613 clinical specimens. Conclusions: The multiplex real-time PCR developed in this study can be used to identify three species of diphtheria-causing bacteria and screen their toxigenicity quickly and accurately. However, in this study, nodiphtheria-causing bacteria other than C. diphtheriae was found in the clinical samples using the PCR or conventional methods. PCR is more sensitive than the conventional methods and can be used as an additional test in diphtheria laboratories.

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Corynebacterium: Molecular Typing and Pathogenesis of Corynebacterium diphtheriae and Zoonotic Diphtheria Toxin-Producing Corynebacterium Species

Vieira

Ramos

Santos

et al. 2022

Molecular Typing in Bacterial Infections, Volume I

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DNA Sequence Analysis of dtxR Gene (Partial) of Corynebacterium diphtheriae Causing Diphtheria in Jawa and Kalimantan Islands, Indonesia

Cited by 6 publications

References 24 publications

Desain Primer untuk Deteksi Gen Diphtheria Toxin Repressor (dtxR) sebagai Biomarker Bakteri Corynebacterium diphtheriae Menggunakan In Silico PCR

Desain Primer untuk Deteksi Gen Diphtheria Toxin Repressor (dtxR) sebagai Biomarker Bakteri Corynebacterium diphtheriae Menggunakan In Silico PCR

Development and Application of Dtxr and Tox Genes Targeting Real-time PCR to Identify Corynebacterium diphtheriae, C. ulcerans, and C. pseudotuberculosis Simultaneously

Corynebacterium: Molecular Typing and Pathogenesis of Corynebacterium diphtheriae and Zoonotic Diphtheria Toxin-Producing Corynebacterium Species

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