An Novel Strategy for Brucella Differential Vaccine Combined with Colloidal Gold Immunochromatographic Strips Based on Mutants of Brucella melitensis M5-90
Aşı sonrası aşılı ve enfekte hayvanları ayırt edebilecek yeni bir aşının geliştirilmesine ihtiyaç vardır. Öncelikli olarak Brucella melitensis'in iki mutantı, sırasıyla 55 ile 152 arası ve 22 ile 185 arası amino asitleri çıkarılmış olan M5-90?bp261 ve M5-90?bp262, üretildi. Sonrasında, her iki mutant da 7 haftalık BALB/c farelere 3.0 × 106 CFU/0.2 mL miktarında inokule edildi. Serum örnekleri Rose Bengal lam testi, serum aglutünasyon testi, VirB5 veya BP26 ile kaplı kolloidal altın immunokromatografik şerit (ICS) ve indirek enzim bağlı immunsorbent assay ile değerlendirildi. Dalak doku örnekleri immunizasyon sonrası 10, 20, 30 ve 40. günlerde B. melitensis varlığını belirlemek amacıyla kullanıldı. Serolojik sonuçlar doğal ve mutant suşların inokulasyon sonrası 10-30. günler arasında zayıf immünolojik reaksiyon oluşturduğunu gösterdi. İnokulasyon sonrası 20-40 günler arasında dalaktan mutant suşların gruplarından doğal suş grubuna oranla daha az koloni oluşturan birim (CFU) belirlendi. ICS B. melitensis'in doğal ve mutant suçlar ile immunize edilmiş farelerin serumlarını ayırt edebildi. Elde edilen sonuçlar, her iki B. melitensis mutant suşun enfekte hayvanlar ile aşılanmış hayvanların ayırt edilmesinde ICS ile kullanıldığında başarılı olabileceğini göstermiştir
Brucella melitensis M5-90 Mutantları Temelli Kolloidal Altın İmmunokromatografik Şerit İle Kombine Brucella Ayırıcı Aşı İçin Yeni Bir Strateji
There is a requirement to develop an novel vaccine to distinguish between vaccinated and infected animals after vaccination. Two mutants of Brucella melitensis M5-90?bp261 and M5-90?bp262, knockout of the fragments amino acids 55-152 and 22-185 respectively, were primarily generated. Then the two mutants were inoculated 7 weeks old BALB/c mice with 3.0 × 106 CFU/0.2 mL parent or mutant strains, Serum samples were evaluated by Rose Bengal plate tests, serum agglutination tests, colloidal gold immunochromatographic strips (ICS) coated with VirB5 or BP26 and indirect enzyme-linked immunosorbent assays. Spleen tissue samples were used to determine B. melitensis abundance at 10, 20, 30, and 40 days post-immunization (dpi). The serological results showed that the parent and mutant strains elicited a weak immunological reaction at 10-30 dpi. Fewer colony forming units (CFUs) were recovered from spleens in mutant strains group than the parent strain group during 20-40 post-immunization (dpi). The ICS were able to distinguish between the sera from mice immunized with either a parent or mutant strain of B. melitensis. These results indicate that the two mutants strain of B. melitensis have potentially perspectives in distinguishing infected animals from vaccination combined with ICS in this study
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