PCR alapú diagnosztikai módszer alkalmazása a sertéspestis kórhatározásában és a járványtani nyomozásban

Translated title of the contribution: PCR as a Diagnostic tool for the diagnosis and epizootological investigation of swine fever

István Kiss, S. Kecskeméti, Endre Bajmócy, J. Tanyi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The economical losses and the difficulties of prevention and control of swine fever demand the most sensitive, specific and the quickest tools for its diagnosis, such as the reverse transcription-polimerase chain reaction (RT-PCR). However, this method alone does not inform about "polymorphism", i.e. the genetic heterogeneity of the amplified RT-PCR product. This can be examined with the single-strand conformational polymorphism (SSCP) method where the double-strand RT-PCR product is run on a polyacrylamide gel after denaturation. This method enables conclusions to be drawn about the "wild" or "mutant" nature of the virus that is represented by the RT-PCR product and the number and complexity of subpopulations in the sample. These methods were used to examine 57 deep frozen samples from previous outbreaks categorised into five groups on the basis of pathology. Directed to the E2 region of the viral genome, RT-PCR quickly and reliably showed evidence of the virus from separated lymphocytes and various organs (most sensitively from the spleen and from tonsils) even after prolonged exposure of the sample to ambient room temperature. Specificity and sensitivity of the RT-PCR was further enhanced by the use of nested primers and a restriction endonuclease (HaeIII). It was shown with SSCP that the RT-PCR products did have different running patterns thus they were assumed to have derived from different subpopulations of the virus. The authors conclude that these techniques can prove useful tools in the prevention and control of infectious diseases.

Original languageHungarian
Pages (from-to)22-28
Number of pages7
JournalMagyar Allatorvosok Lapja
Volume121
Issue number1
Publication statusPublished - 1999

Fingerprint

Classical Swine Fever
reverse transcription
Reverse Transcription
fever
Polymerase Chain Reaction
swine
single-stranded conformational polymorphism
Viruses
viruses
methodology
Genetic Heterogeneity
tonsils
Viral Genome
Palatine Tonsil
DNA Restriction Enzymes
restriction endonucleases
polyacrylamide
denaturation
sampling
infectious diseases

ASJC Scopus subject areas

  • veterinary(all)

Cite this

PCR alapú diagnosztikai módszer alkalmazása a sertéspestis kórhatározásában és a járványtani nyomozásban. / Kiss, István; Kecskeméti, S.; Bajmócy, Endre; Tanyi, J.

In: Magyar Allatorvosok Lapja, Vol. 121, No. 1, 1999, p. 22-28.

Research output: Contribution to journalArticle

@article{a9e94d0d5a01418a9c277effdb549e1e,
title = "PCR alap{\'u} diagnosztikai m{\'o}dszer alkalmaz{\'a}sa a sert{\'e}spestis k{\'o}rhat{\'a}roz{\'a}s{\'a}ban {\'e}s a j{\'a}rv{\'a}nytani nyomoz{\'a}sban",
abstract = "The economical losses and the difficulties of prevention and control of swine fever demand the most sensitive, specific and the quickest tools for its diagnosis, such as the reverse transcription-polimerase chain reaction (RT-PCR). However, this method alone does not inform about {"}polymorphism{"}, i.e. the genetic heterogeneity of the amplified RT-PCR product. This can be examined with the single-strand conformational polymorphism (SSCP) method where the double-strand RT-PCR product is run on a polyacrylamide gel after denaturation. This method enables conclusions to be drawn about the {"}wild{"} or {"}mutant{"} nature of the virus that is represented by the RT-PCR product and the number and complexity of subpopulations in the sample. These methods were used to examine 57 deep frozen samples from previous outbreaks categorised into five groups on the basis of pathology. Directed to the E2 region of the viral genome, RT-PCR quickly and reliably showed evidence of the virus from separated lymphocytes and various organs (most sensitively from the spleen and from tonsils) even after prolonged exposure of the sample to ambient room temperature. Specificity and sensitivity of the RT-PCR was further enhanced by the use of nested primers and a restriction endonuclease (HaeIII). It was shown with SSCP that the RT-PCR products did have different running patterns thus they were assumed to have derived from different subpopulations of the virus. The authors conclude that these techniques can prove useful tools in the prevention and control of infectious diseases.",
author = "Istv{\'a}n Kiss and S. Kecskem{\'e}ti and Endre Bajm{\'o}cy and J. Tanyi",
year = "1999",
language = "Hungarian",
volume = "121",
pages = "22--28",
journal = "Magyar Allatorvosok Lapja",
issn = "0025-004X",
publisher = "Magyar Mezogazdasag Ltd",
number = "1",

}

TY - JOUR

T1 - PCR alapú diagnosztikai módszer alkalmazása a sertéspestis kórhatározásában és a járványtani nyomozásban

AU - Kiss, István

AU - Kecskeméti, S.

AU - Bajmócy, Endre

AU - Tanyi, J.

PY - 1999

Y1 - 1999

N2 - The economical losses and the difficulties of prevention and control of swine fever demand the most sensitive, specific and the quickest tools for its diagnosis, such as the reverse transcription-polimerase chain reaction (RT-PCR). However, this method alone does not inform about "polymorphism", i.e. the genetic heterogeneity of the amplified RT-PCR product. This can be examined with the single-strand conformational polymorphism (SSCP) method where the double-strand RT-PCR product is run on a polyacrylamide gel after denaturation. This method enables conclusions to be drawn about the "wild" or "mutant" nature of the virus that is represented by the RT-PCR product and the number and complexity of subpopulations in the sample. These methods were used to examine 57 deep frozen samples from previous outbreaks categorised into five groups on the basis of pathology. Directed to the E2 region of the viral genome, RT-PCR quickly and reliably showed evidence of the virus from separated lymphocytes and various organs (most sensitively from the spleen and from tonsils) even after prolonged exposure of the sample to ambient room temperature. Specificity and sensitivity of the RT-PCR was further enhanced by the use of nested primers and a restriction endonuclease (HaeIII). It was shown with SSCP that the RT-PCR products did have different running patterns thus they were assumed to have derived from different subpopulations of the virus. The authors conclude that these techniques can prove useful tools in the prevention and control of infectious diseases.

AB - The economical losses and the difficulties of prevention and control of swine fever demand the most sensitive, specific and the quickest tools for its diagnosis, such as the reverse transcription-polimerase chain reaction (RT-PCR). However, this method alone does not inform about "polymorphism", i.e. the genetic heterogeneity of the amplified RT-PCR product. This can be examined with the single-strand conformational polymorphism (SSCP) method where the double-strand RT-PCR product is run on a polyacrylamide gel after denaturation. This method enables conclusions to be drawn about the "wild" or "mutant" nature of the virus that is represented by the RT-PCR product and the number and complexity of subpopulations in the sample. These methods were used to examine 57 deep frozen samples from previous outbreaks categorised into five groups on the basis of pathology. Directed to the E2 region of the viral genome, RT-PCR quickly and reliably showed evidence of the virus from separated lymphocytes and various organs (most sensitively from the spleen and from tonsils) even after prolonged exposure of the sample to ambient room temperature. Specificity and sensitivity of the RT-PCR was further enhanced by the use of nested primers and a restriction endonuclease (HaeIII). It was shown with SSCP that the RT-PCR products did have different running patterns thus they were assumed to have derived from different subpopulations of the virus. The authors conclude that these techniques can prove useful tools in the prevention and control of infectious diseases.

UR - http://www.scopus.com/inward/record.url?scp=0033477123&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033477123&partnerID=8YFLogxK

M3 - Article

VL - 121

SP - 22

EP - 28

JO - Magyar Allatorvosok Lapja

JF - Magyar Allatorvosok Lapja

SN - 0025-004X

IS - 1

ER -