scholarly journals IDENTIFIKASI SUBTIPE Enterotoxigenic Escherichia coli DAN Enteroaggregative Escherichia coli DARI SPESIMEN USAP DUBUR PENJAMAH MAKANAN DI DENPASAR MENGGUNAKAN POLYMERASE CHAIN REACTION

2019 ◽  
Vol 8 (1) ◽  
pp. 7
Author(s):  
D A Indah Gitaswari ◽  
Sri Budayanti
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Shiga Toxin ◽  
Hepatitis A ◽  
Enterohemorrhagic Escherichia Coli ◽  
Enterotoxigenic Escherichia Coli ◽  
Food Handler ◽  
Chain Reaction ◽  
Diarrheagenic Escherichia Coli ◽  
Polymerase Chain

Penjamah makanan atau food handler memiliki risiko paling tinggi untuk terpapar penyakit yang disebabkan oleh bakteri. Penjamah makanan dapat bertindak sebagai carrier penyakit infeksi seperti demam tifoid, hepatitis A, dan diare. Hal ini disebabkan karena kurangnya kemampuan dari penjamah makanan dalam menjaga kebersihan. Bakteri patogen tersering sebagai penyebab diare adalah Escherichia coli yang diperkirakan sebagai penyebab 1.5 juta kematian per tahun. Bakteri Escherichia coli memiliki beberapa subtipe penyebab diare yang disebut Diarrheagenic Escherichia Coli (DEC) diantaranya adalah Enterotoxigenic Escherichia coli (ETEC), Enteropathogenic Escherichia coli (EPEC), Enteroinvasive Escherichia coli (EIEC), Enteroaggregative Escherichia coli (EAEC), Vero toxin-producing/Shiga toxin-producing Escherichia coli (VTEC/STEC), Enterohemorrhagic Escherichia coli (EHEC). Enterotoxigenic Escherichia coli (ETEC) dan Enteroaggregative Escherichia coli (EAEC) paling sering sebagai penyebab diare. Saat ini prevalensi subtipe Escherichia coli sebagai penyebab diare di Bali masih belum diketahui secara pasti. Untuk melihat prevalensi tersebut, pada penelitian ini akan diteliti tipe ETEC dan EAEC dengan menggunakan metode Polymerase Chain Reaction (PCR) unipleks. Sampel merupakan isolat Escherichia coli dari spesimen usap dubur penjamah makanan di Denpasar tahun 2015. Gen target untuk identifikasi ETEC dan EAEC adalah CVD432 (630 bp), LT (273 bp), dan STh (120 bp). Program PCR yang digunakan pada tahap pre denaturasi 95?C, denaturasi 95?C, suhu annealing 55?C, ekstensi 72?C, dan final ekstensi 72?C. Hasil penelitian didapatkan bahwa prevalensi bakteri Escherichia coli menunjukkan 40% memiliki hasil yang positif terhadap gen ETEC atau EAEC. Dari 40% sampel positif, 31,4% merupakan subtipe EAEC; 5,7% subtipe ETEC; dan 3% memiliki kedua gen yaitu ETEC dan EAEC. Kata kunci: Polymerase Chain Reaction, Enteroaggregative Escherichia coli, Enterotoxigenic Escherichia coli

scholarly journals Multiplex Polymerase Chain Reaction Assay for Identification of Enterotoxigenic Escherichia Coli Strains

2001 ◽  
Vol 13 (4) ◽  
pp. 308-311 ◽  
Author(s):  
Jacek Osek
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Direct Determination ◽  
Enteric Bacteria ◽  
Enterotoxigenic Escherichia Coli ◽  
Chain Reaction ◽  
Gram Negative ◽  
E Coli ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.

scholarly journals High Frequency of Diarrheagenic Escherichia coli in HIV-Infected Patients and Patients with Thalassemia in Kerman, Iran

2015 ◽  
Vol 16 (4) ◽  
pp. 353-358 ◽  
Author(s):  
Hesam Alizade ◽  
Hamid Sharifi ◽  
Zahedeh Naderi ◽  
Reza Ghanbarpour ◽  
Mehdi Bamorovat ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Statistical Analysis ◽  
High Frequency ◽  
Multiplex Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Fecal Samples ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Polymerase Chain

This study was conducted on patients with thalassemia and HIV-infected patients to determine the frequency of diarrheagenic Escherichia coli in Kerman, Iran. We analyzed 68 and 49 E coli isolates isolated from healthy fecal samples of patients with thalassemia and HIV-infected patients, respectively. The E coli isolates were studied using a multiplex polymerase chain reaction to identify the enterotoxigenic E coli (ETEC), enterohemorrhagic E coli (EHEC), and enteropathogenic E coli (EPEC) groups. Statistical analysis was carried out to determine the correlation of diarrheagenic E coli between HIV-infected patients and patients with thalassemia using Stata 11.2 software. The frequency of having at least 1 diarrheagenic E coli was more common in patients with thalassemia (67.64%) than in HIV-infected patients (57.14%; P = .25), including ETEC (67.64% versus 57.14%), EHEC (33.82% versus 26.53%), and EPEC (19.11% versus 16.32%). The results of this study indicate that ETEC, EHEC, and EPEC pathotypes are widespread among diarrheagenic E coli isolates in patients with thalassemia and HIV-infected patients.

Detection of enterotoxigenic Escherichia coli in water by polymerase chain reaction amplification and hybridization

1994 ◽  
Vol 40 (4) ◽  
pp. 243-249 ◽  
Author(s):  
Z. Tamanai-Shacoori ◽  
A. Jolivet-Gougeon ◽  
M. Pommepuy ◽  
M. Cormier ◽  
R. R. Colwell
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Amplification ◽  
Pcr Amplification ◽  
Environmental Water ◽  
Latex Agglutination ◽  
Enterotoxigenic Escherichia Coli ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Polymerase Chain

Enterotoxigenic Escherichia coli was studied in waste water, river water, and seawater from six locations along the west coast of Normandy by using the polymerase chain reaction (PCR) to amplify the heat labile (LT) gene. Cellular DNA was extracted from centrifugation pellets and amplified using PCR. The PCR products were detected by gel electrophoresis and confirmed by hybridization assay, using an 850 base pair HindIII DNA fragment probe from pEWD299 conjugated to digoxigenin and specific for the LT gene. Results of the PCR amplification were compared with those of GM1 enzyme-linked immunosorbent assay, latex agglutination, and colony hybridization. The PCR method was found to be more precise and less time consuming, especially when compared with methods requiring culture of isolates for enumeration of enterotoxigenic E. coli in water.Key words: enterotoxigenic Escherichia coli, PCR, environmental water, digoxigenin.

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