Analisis Marka Gen Patogenik iutA Escherichia Coli Penyebab Colibasilosis pada Ayam Buras

2021 ◽  
pp. 57
Author(s):  
Kadek Satria Adi Marhendra ◽  
I Gusti Ngurah Kade Mahardika ◽  
I Nengah Kerta Besung ◽  
I Gusti Ketut Suarjana
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Avian Pathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Pathogenic Escherichia Coli ◽  
Polymerase Chain

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.

scholarly journals Prevalensi Strain Avian Pathogenic Escherichia coli (APEC) Penyebab Kolibasilosis pada Burung Puyuh

2019 ◽  
Vol 37 (1) ◽  
pp. 69
Author(s):  
Wahyu Prihtiyantoro ◽  
Khusnan Khusnan ◽  
Mitra Slipranata ◽  
Imron Rosyidi
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Meat Products ◽  
Processed Meat ◽  
Avian Pathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Air Sacs ◽  
Pathogenic Escherichia Coli ◽  
Polymerase Chain

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. 

scholarly journals Typing of avian pathogenic Escherichia coli strains by REP-PCR

2006 ◽  
Vol 26 (2) ◽  
pp. 69-73 ◽  
Author(s):  
Marcelo Brocchi ◽  
Alessandra Ferreira ◽  
Marcelo Lancellotti ◽  
Eliana G. Stehling ◽  
Tatiana A. Campos ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Clonal Structure ◽  
Avian Pathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Discriminating Power ◽  
Pathogenic Escherichia Coli ◽  
Molecular Sizes ◽  
Polymerase Chain ◽  
Clonal Variability ◽  
Repetitive Extragenic Palindromic

In the present study the repetitive extragenic palindromic (REP) polymerase chain reaction (PCR) technique was used to establish the clonal variability of 49 avian Escherichia coli (APEC) strains isolated from different outbreak cases of septicemia (n=24), swollen head syndrome (n=14) and omphalitis (n=11). Thirty commensal strains isolated from poultry with no signs of these illnesses were used as control strains. The purified DNA of these strains produced electrophoretic profiles ranging from 0 to 15 bands with molecular sizes varying from 100 bp to 6.1 kb, allowing the grouping of the 79 strains into a dendrogram containing 49 REP-types. Although REP-PCR showed good discriminating power it was not able to group the strains either into specific pathogenic classes or to differentiate between pathogenic and non-pathogenic strains. On the contrary, we recently demonstrated that other techniques such as ERIC-PCR and isoenzyme profiles are appropriate to discriminate between commensal and APEC strains and also to group these strains into specific pathogenic classes. In conclusion, REP-PCR seems to be a technique neither efficient nor universal for APEC strains discrimination. However, the population clonal structure obtained with the use of REP-PCR must not be ignored particularly if one takes into account that the APEC pathogenic mechanisms are not completely understood yet.

scholarly journals Virulence-Associated Gene Profiles of Avian Pathogenic Escherichia coli Isolated From Broilers With Colibacillosis: A Pilot Study in Iran

2019 ◽  
Vol 7 (1) ◽  
pp. 4-8
Author(s):  
Payam Haghighi Khoshkhoo ◽  
Hadi Pourtaghi ◽  
Gita Akbariazad ◽  
Saeed Mokhayeri
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Economic Losses ◽  
Avian Pathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Gene Profiles ◽  
Pathogenic Escherichia Coli ◽  
Poultry Farms ◽  
Polymerase Chain

Background: Avian pathogenic Escherichia coli (APEC) causes economic losses in the chicken industry worldwide. Objective: In this study, virulence-associated gene profiles of APEC isolates were investigated by polymerase chain reaction (PCR). Materials and Methods: A total of 60 Escherichia coli isolates were collected from 60 colibacillosis cases from 30 broiler poultry farms in Alborz, Tehran, and Golestan provinces, Iran. After identification by biochemical tests, DNA was extracted by boiling method and 5 virulence-associated genes including: iutA, hlyF, iroN, ompT, and iss were detected by 2 multiplex PCR protocols. Results: Of the 60 APEC isolates, 26 (43.3%) isolates had at least three virulence genes from which 12 (20%) isolates were positive for all 5 virulence genes, whereas 34 (56.6%) carried no investigated virulence genes. Presence of iutA, hlyF, iroN, ompT, and iss genes in the APEC isolates were 17 (28.3%), 17 (28.3%), 24 (40%), 26 (43.3%), and 23 (38.3%), respectively. Conclusion: According to the results, four different virulence-associated gene profiles were seen in isolates, from which profile 1 with 12 (20%) isolates was predominant. These findings were in agreement with the previous reports.

Molecular Analysis of Uropathogenic E.coli Isolates from Urinary Tract Infections

2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Uropathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tract Infections

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.

scholarly journals Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

2007 ◽  
Vol 59 (2) ◽  
pp. 508-512 ◽  
Author(s):  
B.R. Paneto ◽  
R.P. Schocken-Iturrino ◽  
C. Macedo ◽  
E. Santo ◽  
J.M. Marin
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Raw Milk ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Raw Milk Cheese

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

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.

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