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. 

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 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 REAL TIME POLYMERASE CHAIN REACTION (RT-PCR) FOR DNA PIG CONTAMINATION IDENTIFICATION SAUSAGE IN SIX DISTRICT PANDEGLANG BANTEN

2021 ◽  
Vol 1 (1) ◽  
pp. 81-88
Author(s):  
Hadi Susilo
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Meat Product ◽  
Meat Products ◽  
Pcr Analysis ◽  
Processed Meat ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Dna Purity ◽  
Polymerase Chain

Sausage is a meat product processed that is popular food especially in Pandeglang, Banten Province. The importance of halal certificates or the existence of the MUI (Indonesian Ulama Council) halal logo for processed meat products makes Muslim people confident to consume them. The aim this research was to identify pig DNA contamination in sausage products in six  districts in Pandeglang without the MUI halal labels using RT-PCR (Real Time-Polymerase Chain Reaction). RT PCR that can calculate to pig to fill these sample free from pig contamination. This research was divided into two stage, the first stage is extracted or carried out DNA and the second stage is RT PCR analysis. The results of the DNA purity test on sausage samples had DNA purity values ​​of 1.84-1.9 (A260 / A280) and resulted in sample concentrations ranging from 37.8 to 102.5 ng / µl.  The only amplification on the FAM curve was in the positive control pig.  the Cq value ranges from 30 - 31.29. The results of RT PCR on sausage samples in the district area in Pandeglang Banten did not detect the presence of pig DNA.

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.

scholarly journals Development of a Method to Detect Cattle Material from Processed Meat Products Using a Polymerase Chain Reaction

2017 ◽  
Vol 49 (2) ◽  
pp. 135-140
Author(s):  
Young Chul Kwon ◽  
Do-Yun Hah ◽  
Yunwi Heo ◽  
Tae-Kyu Kim ◽  
Yoo-Jeong Choi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Meat Products ◽  
Processed Meat ◽  
Chain Reaction ◽  
Polymerase Chain

Polymerase chain reaction with lateral flow sensor assay for the identification of horse meat in raw and processed meat products

2021 ◽  
Vol 345 ◽  
pp. 128840
Author(s):  
Yang Chen ◽  
Yulong Wang ◽  
Mingya Xiao ◽  
Shuqin Wei ◽  
Haiyan Yang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Meat Products ◽  
Lateral Flow ◽  
Flow Sensor ◽  
Processed Meat ◽  
Chain Reaction ◽  
Horse Meat ◽  
Polymerase Chain

scholarly journals Detection of DNA pork in processed meat products with real-time polymerase chain reaction

Food Research ◽  
2020 ◽  
Vol 4 (5) ◽  
pp. 1719-1725
Author(s):  
W.P. Rahayu ◽  
A.R.W. Dianti ◽  
Nurjanah S. ◽  
N. Pusparini ◽  
W. Adhi
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Melting Curve ◽  
Meat Products ◽  
Processed Meat ◽  
Cyt B ◽  
Chain Reaction ◽  
Ct Value ◽  
Polymerase Chain ◽  
Cyt B Gene

Real-time polymerase chain reaction (qPCR) technique is a suitable method for identifying animal species in processed meat because of its ability to amplify a few fragments of DNA. A specific fragment of pork (mitochondrial cytochrome b gene (cytb)) was used as a DNA ladder. This study aimed to evaluate the use of a cyt-b gene generated primer for detecting the presence of pork in processed meat products by qPCR and determining the threshold cycle cut-off. The evaluation of the primer effectiveness was performed by threshold cycle (Ct) value, amplicon size by electrophoresis and melting curve. Two corned (A, B) and two jerkies (C, D) collected from the market were used as the sample. Genomic DNA from samples, fresh beef (as negative control) and fresh pork (as positive control) were extracted using Qiagen Kits. Amplification condition for 50 cycles of the cyt-b gene was performed as the initial step at 95°C for 10 mins, denaturation step at 95°C for 15 s, annealing step at 55°C for 60 s, extension step at 72°C for 30 s and post-PCR at 72°C for 3 mins. The threshold cycle (Ct) cut-off less than 30 confirmed as pork positive. The result obtained indicated that sample A and D were pork negative, with Ct value respectively 40.73 and 43.59. Melting temperatures of amplicon were ranged from 79.5 to 80.5°C, only differed by 1°C, and the amplicon electrophoresis resulting in a single band of the same size (149 bp). Hence, the melting curve analysis and electrophoresis of PCR products were not able to differentiate between pork and beef.

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.

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