EXTRACTION AND MOLECULAR EVALUATION OF NESISSERIA MENINGITIDES SUBGROUP B OUTER MEMBRANE VESICLE CONTAINING PORA.

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SIADAT S.D.; BEHZADIANNEZHAD GH.; TABARAEI BAHMAN; NAJAR PIRAYEH SH.; AHMADI HOJAT; KAZEMNEJAD ANOUSHIRAVAN; NEJATI MAHDI

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Journal: DANESHVAR MEDICINE Year:2006 | Volume:14 | Issue:65 Page(s): 23-32

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Title

EXTRACTION AND MOLECULAR EVALUATION OF NESISSERIA MENINGITIDES SUBGROUP B OUTER MEMBRANE VESICLE CONTAINING PORA.

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Keywords

OMV-PORAQ3

NEISSERIA MENINGITIDISQ3

SUBGROUP BQ3

IMMUNOGENQ3

EXTRACTIONQ3

Abstract

Introduction: Bacterial meningitis, caused by different groups of Neisseria meningitides, is still one of the serious health problems worldwide. Capsular polysaccharide of group A and C in general, and W135 and Y, in particular, have been known to be a safe protective vaccine for more than two decades. In contrast, the group B capsular polysaccharide was found to be nonimmunogenic in many controlled field trails. For this reason, the SUBGROUP B has become the main causative agent of meningococcal disease in the developing countries. Nowadays, outer membrane protein class I (PorA) has been considered a potential sub-unit IMMUNOGEN to control the disease. Material & methods: In the present investigation, standard group B strain of NEISSERIA MENINGITIDIS (CSBPI, G-245) was cultured under controlled submerge in fermenter containing modified Frantz medium. The cells were harvested at the last exponential growth phase. OMV-PORA was extracted from cell wall through Deoxycholate EXTRACTION Technique and was further purified by sequential centrifugation and ultracentrifugation. The pletted OMV-PORA was homogenized in PBS (pH=7.2), containing sucrose and thiomersal as preservatives. Molecular evaluation of OMV-PORA was done by micrograph scanning under electron microscope and SOS-PAGE. The OMVPorA was identified by Double Diffusion Gel Technique through using hyper immune rabbit's anti OMV-PORA anti sera against purified vesicles.Results: In micrograph scanning of electron microscope, more than 70-90 percent of the OMV-PORA retained its native configurational structure ranging from 50 to 150 nm in size after using EXTRACTION and purification procedures. Purified vesicle showed a strong band of 40-45 KD molecular weights when run on 10% polyacrylamide gel electrophoresis with SOS. A strong precipitate line between hyper immune rabbit's anti OMV-PORA anti sera and purified vesicles was shown by Double Diffusion Gel Technique. Besides, the OMV-PORA was found to be safe when rabbit pyrogenic test and abnormal toxicity test in mice and guinea pigs were performed.Discussion: The obtained results reveal that, OMV-PORA contains intact native structure and composition after EXTRACTION and purification. Thus, OMV-PORA can be regarded as a safe potent IMMUNOGEN to control group B meningococcal disease.

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APA: Copy

SIADAT, S.D., TABARAEI, BAHMAN, AHMADI, HOJAT, BEHZADIANNEZHAD, GH., KAZEMNEJAD, ANOUSHIRAVAN, NAJAR PIRAYEH, SH., & NEJATI, MAHDI. (2006). EXTRACTION AND MOLECULAR EVALUATION OF NESISSERIA MENINGITIDES SUBGROUP B OUTER MEMBRANE VESICLE CONTAINING PORA.. DANESHVAR MEDICINE, 14(65), 23-32. SID. https://sid.ir/paper/30475/en

Vancouver: Copy

SIADAT S.D., TABARAEI BAHMAN, AHMADI HOJAT, BEHZADIANNEZHAD GH., KAZEMNEJAD ANOUSHIRAVAN, NAJAR PIRAYEH SH., NEJATI MAHDI. EXTRACTION AND MOLECULAR EVALUATION OF NESISSERIA MENINGITIDES SUBGROUP B OUTER MEMBRANE VESICLE CONTAINING PORA.. DANESHVAR MEDICINE[Internet]. 2006;14(65):23-32. Available from: https://sid.ir/paper/30475/en

IEEE: Copy

S.D. SIADAT, BAHMAN TABARAEI, HOJAT AHMADI, GH. BEHZADIANNEZHAD, ANOUSHIRAVAN KAZEMNEJAD, SH. NAJAR PIRAYEH, and MAHDI NEJATI, “EXTRACTION AND MOLECULAR EVALUATION OF NESISSERIA MENINGITIDES SUBGROUP B OUTER MEMBRANE VESICLE CONTAINING PORA.,” DANESHVAR MEDICINE, vol. 14, no. 65, pp. 23–32, 2006, [Online]. Available: https://sid.ir/paper/30475/en

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