Zastosowanie metody PCR do wykrywania Listeria monocytogenes w mleku

• Autorzy: Wieckowska M. , Kotlowski R. , Kur J. , Rudnicka W.
• Języki publikacji: PL

Abstrakty

PL

Przeprowadzono elektroforetyczne i serologiczne badania porównawcze składu białek 6 szczepów M. pneumoniae i 1 szczepu M. genitalium. Stwierdzono pełne podobieństwo budowy antygenowej badanych szczepów M. pneumoniae oraz bliskie podobieństwo antygenowe M. genitalium do M. pneumoniae. Najbardziej swoistymi spośród antygenów M. pneumoniae okazały się białka o masie cząsteczkowej 170 (białko P1) i 89 kDa, uważane za najważniejsze adhezyny komórki mykoplazmowej.

EN

The Mycoplasma pneumoniae FH strain routinely used in our laboratory for over 25 years as antigen in serological tests, 2 reference M. pneumoniae strains from ATCC (29342 and M129) and 3 isolates of M. pneumoniae obtained in 1995 from pneumonia patients were compared by SDS-PAGE, complement fixation test (CFT) and by Western-immunoblotting against human and rabbit serum samples with high level of mycoplasmal antibodies. On SDS-PAGE all M. pneumoniae strains showed the same number of 23 polypeptides on the gel with identical molecular weights. The same strains on immunoblotting against human and rabbit serum samples showed six bands: 170, 89, 75, 55, 38 and 33 kDa with the strongest antibody staining in 170-(P1 protein) and 89-kDa bands. Because of its known antigenic relationships Mycoplasma genitalium was used for comparison. The pattern of M genitalium proteins on SDS-PAGE was similar to pattern of M. pneumoniae but distinguishable. On immunoblotting six proteins of M. genitalium (135, 127, 110, 95, 75 and 45 kDa) reacted with human and rabbits immunoglobulins for M. pneumoniae antigens. Furthermore in complement fixation test both antigens, prepared from M. pneumoniae and M. genitalium, reacted as well with human and rabbit immunoglobulins for M. pneumoniae and with rabbit immunoglobulins for M. genitalium. These cross-reactions observed in serological techniques could give false positive results in routine diagnosis of M. pneumoniae infections. In such situations showing on immunoblott of presence in tested serum sample of antibodies to 170 - and 89 kDa proteins could confirm M. pneumoniae infection.

Słowa kluczowe

PL

listerioza; choroby czlowieka; bakterie Gram-dodatnie; Listeria monocytogenes; lancuchowa reakcja polimerazy; badania mikrobiologiczne; mleko; oznaczanie

• Wydawca: -
• Czasopismo: Medycyna Doświadczalna i Mikrobiologia
• Rocznik: 1998
• Tom: 50
• Numer: 3-4
• Strony: 251-257
• Opis fizyczny: s.251-257,fot.,bibliogr.

Twórcy

autor

Wieckowska M.

• Uniwersytet Lodzki, 90-237 Lodz, ul.S.Banacha 12-16

autor

Kotlowski R.

autor

Kur J.

autor

Rudnicka W.

Bibliografia

• 1. Aubert G, Pozzetto B, Hafid J, Gaudin OG. Immunoblotting patterns with Mycoplasma pneumoniae of serum specimens from infected and non-infected subjects. J Med Microbiol 1992; 36: 341-46.
• 2. Bredt W, Kleinmann B, Jacobs E. Antibodies in the sera of Mycoplasma pneumoniae-infected patients against proteins of Mycoplasma genitalium and other mycoplasmas of man. Zbl Bakt Hyg 1987; 266: 32-42.
• 3. Cimolai N, Mah D, Thomas E, Middleton PJ. Rapid immunoblot method for diagnosis of acute Mycoplasma pneumoniae infection. Eur J Clin Microbiol Inf Dis 1990; 9: 223-26.
• 4. Clyde WA, Hu PC. Antigenic determinants of the attachment protein of Mycoplasma pneumoniae shared by other pathogenic Mycoplasma species. Inf Immun 1986; 51: 690-92.
• 5. Dallo SF, Chavoya A, Su CJ, Baseman JB. DNA and protein sequence homologies between the adhesins of Mycoplasma genitalium and Mycoplasma pneumoniae. Inf Immun 1989; 57: 1059-65.
• 6. Hu PC, Schaper U, Colier AM i inni. A Mycoplasma genitalium protein resembling the Mycoplasma pneumoniae attachment protein. Inf Immun 1987; 55: 1126-31.
• 7. Jacobs E, Buchholz A, Kleinmann B, Bredt W. Use of adherence protein of Mycoplasma pneumoniae as antigen for enzyme-linked immunosorbent assay (ELISA). Isr J Sci 1987; 23: 709-12.
• 8. Jacobs E, Fuchte K, Bredt WA 168-kilodalton protein of Mycoplasma pneumoniae used as antigen in a dot enzyme-linked immunosorbent assay.Eur J Clin 1986; 5: 435-40.
• 9. Jacobs E, Waiter T, Schaefer HE, Bredt W. Comparison of host responses after intranasal infection of guinea-pigs with Mycoplasma genitalium or with Mycoplasma pneumoniae. Microb Path 1991; 10: 221-29.
• 10. Kałużewski S. An evaluation of the applicability of some serologic tests in the diagnosis of Mycoplasma pneumoniae infections. I. Antigen for complement fixation test. Exp Med Microbiol 1972; 24: 334-45.
• 11. Kenny GE, Cartwright FD. Immunoblotting for determination of the antigenic specificities of antibodies to the Mycoplasmatales. Isr J Med Scien 1984; 20: 908-11.
• 12. Kleemola M, Kayhty H. Increase in titers of antibodies to Mycoplasma pneumoniae in patients with purulent meningitis. J Infect Dis 1982; 146: 284-88.
• 13. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacte¬riophage T4. Nature 1970; 227: 680-85.
• 14. Leinikki PO, Pantzar P, Tykkä H. Antibody response in patients with acute pancreatitis to Mycoplasma pneumoniae. Scand J Gastroenterol 1973; 8: 631-35.
• 15. Lind K. Serological cross-reactions between Mycoplasma genitalium and Mycoplasma pneu¬moniae. Lancet 1982; ii: 1158-59.
• 16. Lind K, Linhardt B, Schütten HJ i inni. Serological cross-reactions between Mycoplasma genitalium and Mycoplasma pneumoniae. J Clin Microb 1984; 20: 1036-43.
• 17. Pönka A, Pönka T, Sarna S, Penttinen K. Questionable specificity of lipid antigen in the Mycoplasma pneumoniae complement fixation test in patients with extrapulmonary ma¬nifestations. J Infect 1981; 3: 332-38.
• 18. Rastawicki W. Ocena przydatności odczynu western-immunoblotting do badania humoralnej odpowiedzi na antygeny Mycoplasma pneumoniae w przebiegu naturalnego zakażeniu u lu¬dzi. Med Dośw Mikrobiol 1996; 48: 39-48.
• 19. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci 1979; 76: 4350-54.
• 20. Vu AC, Foy HM, Cartwright FD, Kenny GE. The principal protein antigens of isolates of Mycoplasma pneumoniae as measured by levels of immunoglobulin G in human serum are stable in strains collected over a 10-year period. Inf Immun 1987; 55: 1830-36.