You are here

Results of commission trials kit for detecting yersinia enterocolitica dna by polymerase chain reaction

In connection with the signing of the Economic Association Agreement between Ukraine and the European Union, changes in domestic food legislation in order to harmonize it with European, the need of implementation in Ukraine European model of product conformity assessment requirements of technical regulations to veterinary medicine as one of the most important links in obtaining high-quality and safe products possible if animals debugging proper control at all stages of production with identified critical points of possible contamination, including biotic factors.

One of the potentially hazardous representatives of optional (transit) microflora of different biotypes of animal organism is the causative agent of intestinal yersiniosis Yersinia enterocolitica.

According to the literature major source of yersiniosis infection for human are agricultural products - both animal and plant origin. In Yersinia carrier of cattle indicates numerous reports of false-positive reaction for brucellosis during the execution of national programs to combat the disease (including cross-reactions were found to Yersinia enterocolitica serotype O: 9).

The level of contamination yersiniis food according to EFSA ranged from 0 to 83%, and it is most often were contaminated pork tongue (83%), liver (73%), heart (71%) and kidney (67%). Vishnubhatla co-authors found high levels of contamination with minced beef yersiniis. There is evidence of contamination iyersyniyamy of oysters, mussels, shrimp, crab, fish, salads with chicken, stewed mushrooms, cabbage, celery, carrots, etc. [5].

Diagnosis of yersiniosis conducted integrated with the epizootic data, clinical and pathological changes and necessary laboratory tests that are hard and entail a great time. The main diagnostic methods yersiniosis include: 1) bacteriological method, 2) biological method, 3) serological method 4) polymerase chain reaction (PCR).

The most rapid and highly specific method for diagnosis of yersiniosis is polymerase chain reaction (PCR).

Given the diversity of biotypes of bacteria species Yersinia enterocolitica, as well as numerous types that are not cultivated for their detection has been developed test system based on "nested" version of PCR. As the target was selected gene encoding subunit 16S [6].

Test system «Yersinia enterocolitica - PCR Test" conducted by the algorithm:

1. Definition of the exterior components of the test system;

2. Determination of the specificity of the test system;

2.1.1. Preparation for analysis;

2.1.2. Setting PCR;

2.1.3. Еlectrophoresis analysis of PCR products;

2.1.4. Accounting of results;

3. Determination of sensitivity;

4. Determination of the reproducibility of the results.

Benefits of yersiniosis detection by PCR: 1) high specificity, due to the fact that the experimental material showing typical only for Yersinia DNA fragment; 2) high sensitivity, PCR makes it possible to detect DNA of Yersinia biotypes that are not cultivated; 3) high speed of result, the analysis does not require selection or accumulation of the pathogen, the study 1 sample takes an average of 6-8 hours.

The use of laboratory practice modern methods of diagnosis improves the performance of early diagnosis and epidemiological surveillance for yersiniosis.

Key words: Yersinia enterocolitica, polymerase chain reaction, DNA of bacteria, diagnostic kit.

1. Zakon Ukrai'ny pro vnesennja zmin do dejakyh zakonodavchyh aktiv Ukrai'ny shhodo harchovyh produktiv / Verhovna Rada Ukrai'ny. – Ofic. vyd. – K.: Parlam. vyd-vo, 2014. – № 41–42, st. 2024.

2. Yersinia enterocolitica: Epidemiological Studies and Outbreaks [Електроний ресурс] // Journal of Pathogens Vol. 2011/ – Vol 2011 – P. 4, 6  / Спосіб доступу

3. Bakteriologicheskaja i serologicheskaja diagnostika iersiniozov zhivotnyh / Materialy Respublikanskoj konferencii «Veterinarnaja medicina. Jekonomicheskie, social'nye i jekologicheskie problem» (20-22 nojabrja 1990 g.). / A.F. Babkin, V.G. Skripnik, L.B. Ivanovskaja – Har'kov, 1990. – S. 269–270.

4. Rezul'taty gematologicheskih issledovanij pri jeksperimntal'nom iersinioze / Materialy Vsesojuznoj konferencii. Povyshenie produktivnosti s.-h. zhivotnyh i sovershenstvovanie mer bor'by s boleznjami v uslovijah intensivnogo vedenija zhivotnovodstva i sozdanija fermers'kih hozjajstv: (17-22 sentjabrja 1991 g.). / V.G. Skrypnik, L.B. Ivanovskaja //– Har'kov, 1991. – S. 145–146.

5. Monitoring and identification of human enteropathogenic Yersinia spp. [Текст] / The EFSA Journal. – 2007. – Vol. 3.1., 3.2. – P. 10–13.

6. N.M.Sobolev. Serologicheskaja i bakteriologicheskaja indikacija iersinij u zhivotnyh / V. Orlova, V. Skrypnik, E. Savinova, L. Ivanovskaja. // Veterinarija, №63. – K:. Urozhaj, 1988.– S. 20–24.

7. Kozlovs'ka G.V. Kontrol' mikroflory biotopiv tvarynnogo organizmu – vazhlyvyj element v organizacii' otrymannja jakisnoi' j bezpechnoi' produkcii' [Tekst] / G. Kozlovs'ka, V. Skybic'kyj // Visnyk poltavs'koi' agrarnoi' akademii'. – 2013. – №4. – S. 56 – 58.

8. Epidemichna znachymist' ijersynioziv v m. Kyjevi / Materialy narady – seminaru z aktual'nyh pytan' epidemiologii' i profilaktyky zooantroponoznyh infekcijnyh hvorob ta rezhymu bezpeky u laboratorijah derzhavnoi' sanitarno – epidemiologichnoi' sluzhby MOZ. / O.M. Ruban, O.V. Nazarenko, M.A. Kravchenko, V.V. Ljashenko. – Dnipropetrovs'k, 2006. – S.122–123.

9. Elmanama A.A. Yersinia enterocolitica and Aeromonas hydrophila in Clinical, Food and Environmental Samples in Gaza / Abdelraouf A. Elmanama, Naima Ferwana // Journal of Al Azhar University-Gaza (Natural Sciences). – 2011. – № 13. – P. 69-82.

10. Ivanovs'ka L.B. / Pro klinichni oznaky, shho sposterigajut'sja u riznyh vydiv tvaryn hvoryh na ijersynioz / L.B. Ivanovs'ka //Visnyk Sums'kogo NAU, 2003.– №8. – S. 40–43.

11. Zoonoses and public health/ Zur diagnostik und vorkommen von Yersinia enterocolitica in wasser./ [Електронний ресурс] / Спосіб доступу 0450.1986.tb00009 x/abstract