Zastosowanie metody multiplex PCR i PCR-RFLP do identyfikacji Bacillus anthracis

• Autorzy: Szymajda U. , Bartoszcze M.

Warianty tytułu

EN

Application of the multiplex PCR and PCR-RFLP method in the identification of the Bacillus anthracis

• Języki publikacji: PL

Abstrakty

PL

Zastosowano metodę multiplex PCR i PCR-RFLP (PCR-restriction fragment length polymorphism) do identyfikacji szczepów Bacillus anthracis. Metoda multiplex PCR pozwalała na wykrycie genów kodujących czynniki wirulencji lef, cya,pag u szczepów B. anthracis obecnych w plazmidzie pXO1, genu cap występującego w plazmidzie pXO2 oraz markera chromosomalnego Ba813. Analiza genotypowa sekwencji SG-749 z zastosowaniem metody PCR- RFLP wykazała, że wszystkie badane szczepy B. anthracis, charakteryzowały się tym samym wzorem restrykcyjnym złożonym z dwóch fragmentów DNA o wielkości 90 i 660 bp, co umożliwiło odróżnienie ich od innych blisko spokrewnionych gatunków bakterii z grupy B. cereus posiadających różne wzory restrykcyjne.

EN

The aim of this study was to apply the multiplex PCR and PCR-RFLP method for the identification of the B. anthracis strains and to distinguish those bacteria from other members of the Bacillus cereus group. The multiplex PCR method enables to detect the virulence factors, i.e. the toxin and the capsule in B. anthracis strains. To do that, the authors have used 5 primer pairs specific for the fragments of lef, cya,pag genes which are present in the pXO1 plasmid and encode the toxin, the cap gene, which is present in the pXO2 plasmid and encodes the capsule, and the Ba813 chromosomal sequence. Among the four B. anthracis strains examined, three contained two plasmids and the Ba813 chromosomal sequence, while the fourth one contained the pXO1 plasmid only, together and the Ba813 chromosomal sequence. Other bacterial species, belonging to theB. cereus group, were also examined: б strains of B. cereus, 4 strains of B. thuringiensis and one strain ofB. mycoides. The presence of Ba813 chromosomal sequence has been detected in two B. cereus strains. Neither plasmids nor Ba813 chromosomal sequence have been discovered in other B. cereus, B. thuringiensis and B. mycoides strains. The results of the survey indicate that the Ba813 chromosomal sequence does not occur solely in B. anthracis strains. The PCR-RFLP method with the use of SG-749f and SG-749r primers enabled to demonstrate the presence of DNA sequence (SG-749) in B. anthracis, B. cereus, B. thuringiensis and B. mycoides strains. Restriction analysis with enzyme Alul of the SG-749 sequence, has shown the presence of two DNA fragments at the size of about 90 and 660 bp in all B. anthracis strains. The restriction profile obtained was characteristic for B. anthracis strains and it did not occur in other investigated bacterial species belonging to the B. cereus group. It was not observed even in such B. cereus strains in which the presence of Ba813 sequence was discovered and it enabled to differentiate between B. anthracis strains and other closely related species of their, cereus group.

Słowa kluczowe

PL

waglik; czynniki etiologiczne; Bacillus anthracis; identyfikacja; metody analizy; metoda multiplex PCR; metoda PCR-RFLP; markery chromosomowe; analiza genotypowa; plazmid pXO2; fragmentacja DNA

• Wydawca: -
• Czasopismo: Medycyna Doświadczalna i Mikrobiologia
• Rocznik: 2005
• Tom: 57
• Numer: 3
• Opis fizyczny: s.277-285,rys.,tab.,bibliogr.

Twórcy

autor

Szymajda U.

• Wojskowy Instytut Higieny i Epidemiologii, ul.Lubelska 2, 24-100 Pulawy

autor

Bartoszcze M.

Bibliografia

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