scholarly journals Detection of genes encoding ompW and ctxA of Vibrio cholerae isolated from shrimp and shellfish at Kedonganan fish market, Bali-Indonesia

2019 ◽  
Vol 2 (1) ◽  
pp. 1
Author(s):  
Rian Arinta Kusuma Praja ◽  
I Dewa Made Sukrama ◽  
Ni Nengah Dwi Fatmawati
Keyword(s):  
Polymerase Chain Reaction ◽  
Membrane Protein ◽  
Vibrio Cholerae ◽  
Virulence Factors ◽  
Outer Membrane Protein ◽  
Foodborne Disease ◽  
Chain Reaction ◽  
Fish Market ◽  
Polymerase Chain ◽  
Duplex Polymerase Chain Reaction

<p>Contamination of pathogenic bacteria in food can lead to the emergence of foodborne disease. One of foodborne disease which often occurs in some developing countries such as Africa, Southeast Asia, and Latin America is cholera which is caused by <em>Vibrio cholerae</em>. The disease is transmitted through beverages and food, especially contaminated seafood. <em>V. cholerae</em> has several virulence factors including the outer membrane protein W <em>(ompW)</em> and cholerae toxin <em>(ctx).</em>The<em> ompW</em> acts as a protective barrier and can also be used as a marker specific species of <em>V. cholerae</em> and cholerae toxin is an enterotoxin responsible for the incidence of diarrhea in a cholera outbreak produced by pathogenic <em>V. cholerae</em>. This study was an observational study to determine the level of contamination of <em>V. cholerae</em> by detecting the outer membrane protein W <em>(ompW)</em> and cholerae toxin subunit A <em>(ctxA)</em> gene of <em>V. cholerae</em> in shrimp and shellfish sold at Kedonganan fish market. Samples were taken using total sampling technique and obtained 24 samples consisting of 14 shrimp samples and 10 shellfish samples. Samples were examined using culture methods and biochemical tests, and then further tested using Duplex Polymerase Chain Reaction (dPCR) to detect <em>ompW</em> and <em>ctxA</em> gene. The dPCR assay results showed 8 out of 14 (57.1%) samples from shrimp and 1 out of 10 (10%) samples from the shellfish positive carried <em>ompW</em> gene, and found no positive samples carrying the <em>ctxA</em> gene in samples derived from shrimp and shellfish. Chi square test analysis results indicated contamination of <em>V. cholerae</em> in shrimp was higher than shellfish based on <em>ompW</em> gene (p&lt;0.05). It can be concluded that the shrimp and shellfish at Kedonganan fish market are contaminated by <em>V. cholerae</em>. Further research is needed to detect the virulence factors besides <em>ompW</em> and <em>ctxA </em>of<em> V. cholerae</em> in seafood.</p><pre><strong> </strong></pre>Keywords: Foodborne disease, <em>Vibrio cholerae</em>, <em>ompW</em> gene<em>,</em> <em>ctxA</em> gene, and Duplex Polymerase Chain Reaction (dPCR).

scholarly journals Polymerase chain reaction and an outer membrane protein gene probe for the detection ofPorphyromonas gingivalis

1996 ◽  
Vol 138 (2-3) ◽  
pp. 167-172 ◽  
Author(s):  
Koichi Hiratsuka ◽  
Wataru Yoshida ◽  
Mitsuo Hayakawa ◽  
Hisashi Takiguchi ◽  
Yoshimitsu Abiko
Keyword(s):  
Polymerase Chain Reaction ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Protein Gene ◽  
Gene Probe ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Membrane Protein Gene

scholarly journals Molecular characterization of Candida dubliniensis and Candida albicans in the oral cavity of drug abusers using duplex polymerase chain reaction

2018 ◽  
Vol 4 (1) ◽  
Author(s):  
Parastoo Hassani Abharian ◽  
Parvin Dehghan ◽  
Peyman Hassani Abharian ◽  
Sepideh Tolouei
Keyword(s):  
Polymerase Chain Reaction ◽  
Candida Albicans ◽  
Oral Cavity ◽  
Candida Species ◽  
Species Complex ◽  
Candida Dubliniensis ◽  
Drug Abusers ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Duplex Polymerase Chain Reaction

  Background and Purpose: Candida dubliniensis is closely related to the most pathogenic and prevalent yeast, namely C. albicans. Candida species can opportunistically overgrow in vulnerable individuals and cause a variety of diseases. The current study aimed to identify and isolate C. dubliniensis species present in the Candida albicans species complex identified in the oral cavity of drug abusers. Materials and Methods: This study was conducted on 53 strains of C. albicans species complex, isolated from the oral mucosa of drug abusers in Isfahan, Iran. DNA extraction was accomplished through boiling procedure. Duplex polymerase chain reaction (PCR) was performed to amplify ITS1-5.8S-ITS2 region using four specific primers. Fungal species were identified based on the difference in the size of the bands created in the Agarose gel. Results: Out of the 53 isolates under study, 30 (56.6%) and 14 (26.4%) samples were identified as C. albicans and C. dubliniensis, respectively. In the remaining 9 samples (17%), both types of Candida species were confirmed. Conclusion: The findings of the present study revealed the presence of a noticeable amount of C. dubliniensis in the oral cavity of drug abusers. Therefore, the probable presence of this fungus should be considered during the examination of oral infection among this group. To date, no research has directly investigated this issue in Iran.

Polymerase chain reaction for detection of the cholera enterotoxin operon of Vibrio cholerae.

1991 ◽  
Vol 29 (11) ◽  
pp. 2517-2521 ◽  
Author(s):  
H Shirai ◽  
M Nishibuchi ◽  
T Ramamurthy ◽  
S K Bhattacharya ◽  
S C Pal ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Chain Reaction ◽  
Cholera Enterotoxin ◽  
Polymerase Chain

scholarly journals Rapid Detection of the Vibrio cholerae ctx Gene in Food Enrichments Using Real-Time Polymerase Chain Reaction

2007 ◽  
Vol 90 (5) ◽  
pp. 1278-1283 ◽  
Author(s):  
Willis Fedio ◽  
George M Blackstone ◽  
Lynne Kikuta-Oshima ◽  
Chitra Wendakoon ◽  
Timothy H McGrath ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Real Time ◽  
Bottled Water ◽  
Qpcr Assay ◽  
Second Phase ◽  
Chain Reaction ◽  
Isolation Frequency ◽  
Polymerase Chain ◽  
Static Conditions

Abstract A real-time polymerase chain reaction (qPCR) assay for the detection of the ctxA gene of toxigenic Vibrio cholerae (Vc) was validated against standard culture techniques. The first experimental phase determined optimal enrichment conditions for detection by culture and qPCR of Vc in shrimp, bottled water, milk, and potato salad. The conditions tested included temperature (35 and 42C), time (6 and 18 h), and effect of shaking (0 and 100 rpm). No definitive trends were found with enrichment temperature or shaking on Vc isolation frequency or detection by qPCR. Generally, Vc was detected by qPCR more frequently than Vc was isolated, but this difference was significant only in the 35C 6 h enrichment without shaking. In the second phase of experiments, shrimp, bottled water, milk, potato salad, and oysters were inoculated with each of 3 toxigenic Vc strains (Latin American O1 strain, an O139 strain, and an O1 strain from the U.S. Gulf Coast) and enriched under static conditions at 42C for 6 and 18 h. Overall, detection frequency of ctx by qPCR was 98 (88/90) and 100 (90/90) after 6 and 18 h enrichments, respectively, while Vc isolation frequency was 87 (78/90) and 83 (75/90) after 6 and 18 h, respectively. Toxigenic Vc can be detected by qPCR within an 8 h work day using the 6 h enrichment procedure, assuming an initial level of at least 12 colony-forming units/g; however, overnight enrichment may be necessary to detect lower levels. These data indicate that the qPCR assay for ctx is a more reliable, sensitive, and rapid alternative to standard Vc culture methods and is applicable to diverse food products.

scholarly journals Etiology of Diarrhea Requiring Hospitalization in Bangladesh by Quantitative Polymerase Chain Reaction, 2014–2018

2020 ◽  
Author(s):  
Mami Taniuchi ◽  
Kamrul Islam ◽  
Md Abu Sayeed ◽  
James A Platts-Mills ◽  
Md Taufiqul Islam ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Coefficient Of Variation ◽  
Quantitative Polymerase Chain Reaction ◽  
Age Groups ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Surveillance Systems ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background Diarrhea remains a major public health problem and characterization of its etiology is needed to prioritize interventions. However, most data are from single-site studies of children. We tested samples from participants of any age from 11 geographically diverse hospitals in Bangladesh to describe pathogen-specific burdens of diarrhea. Methods We utilized 2 existing diarrhea surveillance systems: a Nationwide network at 10 sentinel hospitals and at the icddr,b hospital. We tested stools from enrolled participants and nondiarrheal controls for enteropathogens using quantitative polymerase chain reaction and calculated pathogen-specific attributable fractions (AFs) of diarrhea. Results We analyzed 5516 patients with diarrhea and 735 controls. Overall, rotavirus had the highest attributable burden of diarrhea (Nationwide AF, 17.7%; 95% confidence interval [CI], 14.3–20.9%; icddr,b AF, 39.9%; 38.0–41.8%), followed by adenovirus 40/41 (Nationwide AF, 17.9%; 95% CI: 13.9–21.9%; icddr,b AF, 16.6%; 95% CI, 14.4–19.4%) and Vibrio cholerae (Nationwide AF, 10.2%; 95% CI, 9.1–11.3%; icddr,b AF, 13.3%; 95% CI: 11.9–15.1%). Rotavirus was the leading pathogen in children &lt;5 years and was consistent across the sites (coefficient of variation = 56.3%). Adenovirus 40/41 was the second leading pathogen in both children and adults. Vibrio cholerae was the leading pathogen in individuals &gt;5 years old, but was more geographically variable (coefficient of variation = 71.5%). Other attributable pathogens included astrovirus, norovirus, Shigella, Salmonella, ETEC, sapovirus, and typical EPEC. Conclusions Rotavirus, adenovirus 40/41, and V. cholerae were the leading etiologies of infectious diarrhea requiring hospitalization in Bangladesh. Other pathogens were important in certain age groups or sites.

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