A new modified medium for Simultaneous Cystinase  and elek tests of bacteria causing diphtheria

Wang, Li; Sunarno, Sunarno; Syarif, Armaji Kamaludi; Karyana, Muhammad; Rosana, Yeva; Moehario, Lucky Hartati

doi:10.15562/bmj.v8i1.1231

Cited by 5 publications

(3 citation statements)

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“…On the other hand, the clinical specimens were identified using conventional methods according to the WHO guidelines for diphtheria laboratory (15). The diphtheria-causing bacteria were identified using API Coryne ® (bioMérieux), while the bacterial toxigenicity was determined using modified Elek tests (16). After the clinical specimens were examined using the conventional methods, the samples were transferred to 0.5 mL Aquadest in the microtube for the DNA extraction.…”

Section: Conventional Methodsmentioning

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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Section: Conventional Methodsmentioning

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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“…A thinner medium with less density and a closer distance between the antitoxin disc and inoculum reduced the time to obtain the results of the test to within 16 to 24 h. Moreover, the authors suggested that extending the incubation time may result in the occurrence of non-specific lines of precipitation. Further modifications proposed in the scientific literature concerned the medium used for the test, for example, the application of Columbia blood agar base instead of Elek base medium [ 41 ] and the application of a modified medium for simultaneous Cystinase and Elek tests [ 42 ].…”

Section: Immunological Assaysmentioning

confidence: 99%

Challenges of Diphtheria Toxin Detection

Prygiel,

Mosiej,

Polak

et al. 2024

Toxins

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Diphtheria toxin (DT) is the main virulence factor of Corynebacterium diphtheriae, C. ulcerans and C. pseudotuberculosis. Moreover, new Corynebacterium species with the potential to produce diphtheria toxin have also been described. Therefore, the detection of the toxin is the most important test in the microbiological diagnosis of diphtheria and other corynebacteria infections. Since the first demonstration in 1888 that DT is a major virulence factor of C. diphtheriae, responsible for the systemic manifestation of the disease, various methods for DT detection have been developed, but the diagnostic usefulness of most of them has not been confirmed on a sufficiently large group of samples. Despite substantial progress in the science and diagnostics of infectious diseases, the Elek test is still the basic recommended diagnostic test for DT detection. The challenge here is the poor availability of an antitoxin and declining experience even in reference laboratories due to the low prevalence of diphtheria in developed countries. However, recent and very promising assays have been developed with the potential for use as rapid point-of-care testing (POCT), such as ICS and LFIA for toxin detection, LAMP for tox gene detection, and biosensors for both.

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“…Exactly 120 mL of distilled water was measured, poured into the flasks containing samples and stirred to soak completely. The level of the mixture was marked, then sample were autoclaved at 121˚C for 15 minutes [12]. Distilled water was added to the initial mark and cooled to 40˚C.…”

Section: Preparation Of Samplesmentioning

confidence: 99%

Investigation of Novel Plant Maerua Shrub (<i>Maerua subcordata</i>) for Cheap and Efficient Bioethanol Production in Kenya

Kangor¹,

Kiplagat²,

Lutta³

et al. 2021

JACEN

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Bioethanol is an attractive source of energy when compared to fossil fuel. It is renewable and environmentally friendly due to its low toxicity and biodegradability. The first generation bioethanol derived from is limited by the high cost of production of these crops and danger posed to food security. This study investigated the use of wild maerua shrub in production of bioethanol in comparison to cultivated food crops. Fermentation was done using Yeasts Y1, Y2 and Y3. Yeasts Y1 and Y2 were isolated from finger millet malt, while Y3 was the commercial yeast Saccharomyces cerevisiae. Fermented plant samples were distilled, oxidized and analysed at 595nm using UV-Visible spectrophotometer. Statgraphics centurion XVI.I was used for statistical analyses. The concentration (g/L) was obtained from a formula and converted to (g/L).The ethanol concentration (g/L) and productivity (g/L/h) were as follows; cassava (64.052 ± 0.098; 1.334), maize (66.670 ± 0.227; 1.389), sorghum (62.382 ± 2.148b; 1.300) and maerua shrub (61.988 ± 0.160, 1.291) which were significantly higher compared to sugarcane molasses (49.978 g/L, 1.041) when fermented by Y2. Mean ethanol concentration (g/L) and productivity (g/L/h) for plants across all yeasts were comparable (p-value = 0.4239). Maerua Shrub should be used as an alternative sugar source for bioethanol production.

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A new modified medium for Simultaneous Cystinase and elek tests of bacteria causing diphtheria

Cited by 5 publications

References 0 publications

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

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

Challenges of Diphtheria Toxin Detection

Investigation of Novel Plant Maerua Shrub (<i>Maerua subcordata</i>) for Cheap and Efficient Bioethanol Production in Kenya

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A new modified medium for Simultaneous Cystinase and elek tests of bacteria causing diphtheria

Cited by 5 publications

References 0 publications

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

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

Challenges of Diphtheria Toxin Detection

Investigation of Novel Plant Maerua Shrub (&lt;i&gt;Maerua subcordata&lt;/i&gt;) for Cheap and Efficient Bioethanol Production in Kenya

Contact Info

Product

Resources

About

Investigation of Novel Plant Maerua Shrub (<i>Maerua subcordata</i>) for Cheap and Efficient Bioethanol Production in Kenya