Rapid Detection of Virulence-Associated Genes in Avian Pathogenic Escherichia coli by Multiplex Polymerase Chain Reaction

2005 ◽  
Vol 49 (2) ◽  
pp. 269-273 ◽  
Author(s):  
Christa Ewers ◽  
Traute Janßen ◽  
Sabine Kießling ◽  
Hans-C. Philipp ◽  
Lothar H. Wieler
Author(s):  
Kadek Satria Adi Marhendra ◽  
I Gusti Ngurah Kade Mahardika ◽  
I Nengah Kerta Besung ◽  
I Gusti Ketut Suarjana

Kolibasilosis merupakan penyakit menular pada ayam yang disebabkan oleh Avian Pathogenic Escherichia coli (APEC). Kemampuan APEC untuk menyebabkan penyakit tergantung pada banyak faktor patogen, salah satunya adalah gen patogenik iutA. Penelitian bertujuan untuk mengetahui sekuen DNA gen iutA APEC di Bali serta kekerabatannya dengan gen iutA dari negara lain. Penelitian ini menggunakan dua isolat APEC asal ayam buras di Kabupaten Tabanan dan Badung yang telah dimurnikan dan tersedia di Laboratorium Bakteriologi Fakultas Kedokteran Hewan, Universitas Udayana. DNA isolat diisolasi dengan Chelex 10%. Produk polymerase chain reaction (PCR) disekuensing di First Base Laboratories Malaysia dengan metode Sanger’s dideoxy nucleotide termination. Kedua hasil sekuen gen iutA memiliki homologi 100% dengan panjang 250 bp. Analisis filogenik dengan 58 data gen iutA di Escherichia coli dan bakteri lain di dunia memiliki 43 situs polimorfik pada tingkat asam nukleat dan 13 di tingkat asam amino. Gen iutA asal Bali berada di dalam satu kelompok dengan gen iutA asal Brazilia (KP657535) tahun 2011, Jerman (LT599825) tahun 2016, dan Cina (CP033635) tahun 2016. Gen ini dapat digunakan sebagai marker patogenik APEC di Indonesia.


2019 ◽  
Vol 37 (1) ◽  
pp. 69
Author(s):  
Wahyu Prihtiyantoro ◽  
Khusnan Khusnan ◽  
Mitra Slipranata ◽  
Imron Rosyidi

Avian Pathogenic Escherichia coli (APEC) is a pathogen that causes colibacillosis in poultry, including salpingitis, omphalitis, cellulitis, swollen head syndrome, coligranuloma yolk sac inflammation, and air sacs inflammation. APEC is a zoonotic strain which spread through raw meat and processed meat products of animals and birds. In this research, the isolation and identification of Escherichia coli were done by using selective media MacConkey, Kligger Iron Agar, and Gram staining. Polymerase chain reaction (PCR) was used to analyse genetopically to detect 16SrRNA genes, vt1 genes, and vt2 genes. Thirty one (55,36%) isolates of 56 specimens collected from quail were detected as Escherichia coli. The detection of APEC strains towards 31 Escherichia coli isolates were done by using polymerase chain reaction (PCR) with vt1 and vt2 specific primer. The results showed that 32,26% (10/31) was APEC strains and 67.74% was non-APEC strains. From 10 isolates, 90% had vt1 gene and 10% had vt2 gene. Escherichia coli isolates were found in eyes (32,26%), infraorbital sinus fluid (32,26%), nasal fluid (16,20%), also in lungs, air sacs, ascites, and heart for 3,2% each. The isolates could not be found in the specimens from the skull. As a zoonotic agent, the isolates have an impact on human health. 


2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.


2001 ◽  
Vol 13 (4) ◽  
pp. 308-311 ◽  
Author(s):  
Jacek Osek

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.


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