Optimisation and comparison of three PCR procedures for molecular identification of Taenia solium

• Autorzy: Karamon J. , Sroka J. , Cecnek T. , Rozycki M. , Chmurzynska E. , Bilska-Zajac E. , Zdybel J. , Nowak P. , Kedzierska J. , Debiak P.
• Języki publikacji: EN

Abstrakty

EN

The aim of the study was to optimise selected PCR methods for identification of T. solium, and to compare their effectiveness and usefulness. The investigation concerned three PCR methods described earlier: PCR I (specific to oncosphere- specific protein Tso31 gene), PCR II (specific to large subunit rRNA gene), and PCR III (cytochrome c oxidases ubunit 1 gene). Each of them needed optimisation in connection with some changes in the procedures. Among the examined procedures, PCR I was found to be the most useful, requiring the least corrections during optimisation - only a higher concentration of polymerase was necessary. Testing an optimised PCR II method showed strong unspecific reactions with E. granulosus and T. saginata. This method was not considered diagnostically useful in distinguishing 7. solium. PCR III method yielded products only when annealing temperature was lowered by 2°C. Under such conditions, there were no unspecific reactions with three others Taenidae parasites; however, annealing at a temperature only 1°C lower generated a distinct unspecific PCR product from T. saginata DNA. Therefore, this method was of limited usefulness. Comparison of the effectiveness of the two selected methods (PCR I and III) in detection of T. solium in successive DNA dilutions showed a large difference between them: in the same DNA sample, PCR I showed positive results in a sample diluted 1:3200, while PCR III failed at dilutions greater than 1:50. The results showed that among the three different methods used in the investigations, the most specific and effective for identification of 7. solium was PCR I.

• Wydawca: -
• Czasopismo: Bulletin of the Veterinary Institute in Puławy
• Rocznik: 2013
• Tom: 57
• Numer: 4
• Opis fizyczny: p.507-512,fig.,ref.

Twórcy

autor

Karamon J.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Sroka J.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Cecnek T.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Rozycki M.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Chmurzynska E.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Bilska-Zajac E.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Zdybel J.

• Department of Parasitology and Invasive Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Nowak P.

• Laboratory of Parasitology, Department of Microbiology, University Hospital of Krakow, 31-501 Krakow, Poland

autor

Kedzierska J.

• Laboratory of Parasitology, Department of Microbiology, University Hospital of Krakow, 31-501 Krakow, Poland

autor

Debiak P.

• Laboratory of Radiology and Ultrasonography, Faculty of Veterinary Medicine, University of Life Sciences, 20-950 Lublin, Poland

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Identyfikatory

DOI

10.2478/bvip-2013-0088