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Isolation and characterization of Borrelia burgdorferi strains from Ixodes ricinus ticks in the southern England.

Sorouri R, Ramazani A, Karami A, Ranjbar R, Guy EC - Bioimpacts (2015)

Bottom Line: The availability of these two isolates enabled their antigenic characterization with SDS-PAGE and western blotting and comparison with two standard isolates.These studies identified six protein antigens with molecular weights of 18, 30, 39, 47, 60 and 88 kDa with particular promise for detecting specific immune responses to B. burgdorferi infection including Lyme disease.As a result of this study, antigenic differences have been seen between the UK isolates and the foreign isolates used as laboratory standards.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.

ABSTRACT

Introduction: Lyme disease is a bacterial infection caused by the spiral-shaped bacterium Borrelia burgdorferi. We investigated the presence and prevalence of Borrelia species in ticks from the southern England.

Methods: One hundred fifty-five cases (103 adult and 52 nymphal ticks) were collected from animal carcases. The midguts were removed, cultured in Barbour/Stoenner/Kelly II (BSK-II) and Barbour/ Stoenner/Kelly F (BSK-F) media and examined by IF, dark-field microscopy, and nested PCR.

Results: From a total 155 cultured ticks, two showed evidence of spirochetes and denoted as SO-1 and SO-2 strains. The availability of these two isolates enabled their antigenic characterization with SDS-PAGE and western blotting and comparison with two standard isolates. These studies identified six protein antigens with molecular weights of 18, 30, 39, 47, 60 and 88 kDa with particular promise for detecting specific immune responses to B. burgdorferi infection including Lyme disease. We also investigated the effect of repeated subculture on the antigenic pattern of UK isolate of B. burgdorferi.

Conclusion: As a result of this study, antigenic differences have been seen between the UK isolates and the foreign isolates used as laboratory standards.

No MeSH data available.


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Mentions: SDS-PAGE and western blotting were carried out on a number of the serial subcultures. The SDS-PAGE profiles and western blotting in one of the sera tested against the isolates have been shown in Figs. 4 and 5, respectively. In SDS-PAGE profile the major differences which occurred could be detected as early as passage 3 (P3), and this also appeared similar to the passage 2 shown in Fig. 4. Passage 1 clearly showed the presence of the putative OSP-B protein at 32k Da; this was weakly detectable at passage 3 and further decreased up to passage 10. However at passage 20, a new band appeared at 33 kDa which appeared to shift to 34 kDa by passage 32. In contrast, P1 lacked at 21/22 kDa band which was observed in all other subcultures. Less marked differences were seen with the 41 kDa band which showed increased intensity after P1 while the 45 kDa showed a progressive decrease in intensity with repeated subculture. In western blotting as can be seen in Fig. 5 the intensity of most bands did not change with subculture and the 32 kDa putative OSP-B which was strongly present only in P1 did not react with the serum at all. In contrast some additional bands not present in P1, in particular a band at 36 kDa, was detected in the subsequent subcultures. Thus maximum detection of antibodies could be achieved not with P1, but with the subsequent subcultures.


Isolation and characterization of Borrelia burgdorferi strains from Ixodes ricinus ticks in the southern England.

Sorouri R, Ramazani A, Karami A, Ranjbar R, Guy EC - Bioimpacts (2015)

© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492187&req=5

Mentions: SDS-PAGE and western blotting were carried out on a number of the serial subcultures. The SDS-PAGE profiles and western blotting in one of the sera tested against the isolates have been shown in Figs. 4 and 5, respectively. In SDS-PAGE profile the major differences which occurred could be detected as early as passage 3 (P3), and this also appeared similar to the passage 2 shown in Fig. 4. Passage 1 clearly showed the presence of the putative OSP-B protein at 32k Da; this was weakly detectable at passage 3 and further decreased up to passage 10. However at passage 20, a new band appeared at 33 kDa which appeared to shift to 34 kDa by passage 32. In contrast, P1 lacked at 21/22 kDa band which was observed in all other subcultures. Less marked differences were seen with the 41 kDa band which showed increased intensity after P1 while the 45 kDa showed a progressive decrease in intensity with repeated subculture. In western blotting as can be seen in Fig. 5 the intensity of most bands did not change with subculture and the 32 kDa putative OSP-B which was strongly present only in P1 did not react with the serum at all. In contrast some additional bands not present in P1, in particular a band at 36 kDa, was detected in the subsequent subcultures. Thus maximum detection of antibodies could be achieved not with P1, but with the subsequent subcultures.

Bottom Line: The availability of these two isolates enabled their antigenic characterization with SDS-PAGE and western blotting and comparison with two standard isolates.These studies identified six protein antigens with molecular weights of 18, 30, 39, 47, 60 and 88 kDa with particular promise for detecting specific immune responses to B. burgdorferi infection including Lyme disease.As a result of this study, antigenic differences have been seen between the UK isolates and the foreign isolates used as laboratory standards.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.

ABSTRACT

Introduction: Lyme disease is a bacterial infection caused by the spiral-shaped bacterium Borrelia burgdorferi. We investigated the presence and prevalence of Borrelia species in ticks from the southern England.

Methods: One hundred fifty-five cases (103 adult and 52 nymphal ticks) were collected from animal carcases. The midguts were removed, cultured in Barbour/Stoenner/Kelly II (BSK-II) and Barbour/ Stoenner/Kelly F (BSK-F) media and examined by IF, dark-field microscopy, and nested PCR.

Results: From a total 155 cultured ticks, two showed evidence of spirochetes and denoted as SO-1 and SO-2 strains. The availability of these two isolates enabled their antigenic characterization with SDS-PAGE and western blotting and comparison with two standard isolates. These studies identified six protein antigens with molecular weights of 18, 30, 39, 47, 60 and 88 kDa with particular promise for detecting specific immune responses to B. burgdorferi infection including Lyme disease. We also investigated the effect of repeated subculture on the antigenic pattern of UK isolate of B. burgdorferi.

Conclusion: As a result of this study, antigenic differences have been seen between the UK isolates and the foreign isolates used as laboratory standards.

No MeSH data available.


Related in: MedlinePlus