Purification and biological analysis of specific antigens (ESAT۶/CFP۱۰) from Mycobacterium tuberculosis

سال انتشار: 1399
نوع سند: مقاله ژورنالی
زبان: انگلیسی
مشاهده: 331

فایل این مقاله در 10 صفحه با فرمت PDF قابل دریافت می باشد

استخراج به نرم افزارهای پژوهشی:

لینک ثابت به این مقاله:

شناسه ملی سند علمی:

JR_IJVST-12-2_008

تاریخ نمایه سازی: 21 اردیبهشت 1400

چکیده مقاله:

The pathogenesis of Mycobacterium tuberculosis (Mtb) is related to its low molecular weight proteins mainly ESAT۶ and CFP۱۰ that are highly specific and potentially useful for the diagnosis of tuberculosis. This research focused on isolation, purification, and characterization of low molecular weight proteins from Mtb. Cultures of Mtb were inactivated by heating at ۶۸ °C for ۹۰ min and ۱۰۰ °C for ۳ hrs, respectively. Inactivated cultures were filtered and the proteins in the supernatant fluid precipitated with two rounds of ammonium sulfate, at ۴ °C. The collected precipitates were dialyzed and subjected to gel chromatography (G-۵۰) and the obtained fractions were analyzed for protein concentrations and molecular weight. ESAT۶ and CFP۱۰ protein complex in the purified fraction was confirmed by Western blotting. Guinea pig sensitization assay was used for estimating the potency of the purified fraction compared to the standard PPD. The maximum amount of low molecular weight proteins were precipitated by ۲۰% ammonium sulfate. SDS-PAGE analysis revealed protein bands of approximately ۱۰-۱۵ kDa. The purity of the proteins was ≥۹۵%, as confirmed by SDS-PAGE. The presence of the ESAT-۶/CFP۱۰ complex was confirmed by Western blot analysis. The purified fractions showed no cross-reaction with BCG or M. avium strain. ESAT-۶/CFP-۱۰ purified by the ammonium sulfate method appeared to be suitable for the development of a diagnostic kit for the detection of Mtb.

نویسندگان

ناهید مژگانی

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

مهدی بابائی

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

نفیسه شکیبامهر

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

محمد محمدطاهری

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

نادر مصوری

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

آرام قائم پناه

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

کیومرث سلیمانی بابادی

Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran.

مراجع و منابع این مقاله:

لیست زیر مراجع و منابع استفاده شده در این مقاله را نمایش می دهد. این مراجع به صورت کاملا ماشینی و بر اساس هوش مصنوعی استخراج شده اند و لذا ممکن است دارای اشکالاتی باشند که به مرور زمان دقت استخراج این محتوا افزایش می یابد. مراجعی که مقالات مربوط به آنها در سیویلیکا نمایه شده و پیدا شده اند، به خود مقاله لینک شده اند :
  • 1. Mattow J, Jungblut PR, Müller EC, Kaufmann SH. Identification ...
  • 2. Elston JW, Thaker HK. Co-infection with human immunodeficiency virus ...
  • 3. Seibert FB, Glen JT. PPD-S was comprised of approximately ...
  • 4. Behr MA, Wilson MA, Gill WP, Salamon H, Schoolnik ...
  • 5. Huebner RE, Schein MF, Bass JB. The tuberculin skin ...
  • 6. Farhat M, Greenaway C, Pai M, Menzies D. False-positive ...
  • 7. Shingadia D, Novelli V. The tuberculin skin test: a ...
  • 8. Coler RN, Skeiky YA, Ovendale PJ, Vedvick TS, Gervassi ...
  • 9. Kumar GSS, Venugopal AK, Selvan LDN, Marimuthu A, Keerthikumar ...
  • 10. Brodin P, Majlessi L, Marsollier L, De Jonge MI, ...
  • 11. Meher AK, Bal NC, Chary KV, Arora A. Mycobacterium ...
  • 12. Alito A, McNair J, Girvin RM, Zumarraga M, Bigi ...
  • 13. Lewinsohn DM, Grotzke JE, Heinzel AS, Zhu L, Ovendale ...
  • 14. Sørensen AL, Nagai S, Houen, G, Andersen P, Andersen ...
  • 15. Vordermeier HM, Whelan A, Cockle PJ, Farrant L, Palmer ...
  • 16. Babaie M, Ghaempanah A, Mehrabi Z, Mollaei A. Partial ...
  • 17. Babaie M, Zolfagharian H, Jamili S, Zolfaghari M. Biochemical, ...
  • 18. Chevallet M, Luche S, Rabilloud T. Silver staining of ...
  • 19. Babaie M, Zolfagharian H, Salmanizadeh H, Mirakabadi AZ, Alizadeh ...
  • 20. Franco J, Camarena J, Nogueira J, Blanquer R, Ruiz ...
  • 21. Weldingh K, Andersen P. ESAT-6/CFP10 Skin Test Predicts Disease ...
  • 22. Mahmoudi S, Mamishi S, Ghazi M, Sadeghi RH, Pourakbari ...
  • 23. Van Pinxteren LA, Ravn P, Agger EM, Pollock J, ...
  • 24. Andersen P, Andersen AB, Sørensen AL, Nagai S. Recall ...
  • 25. Long ER, Seibert FB. The chemical composition of the ...
  • 26. Seibert FB, Pedersen KO, Tiselius A. Molecular weight, electro ...
  • 27. Landi S, Held HR. Physicochemical and biological studies on ...
  • 28. Orme IM, McMurray DN, Belisle JT. Tuberculosis vaccine development: ...
  • 29. Behar SM, Dascher CC, Grusby MJ, Wang CR, Brenner ...
  • 30. Thaveekarn W, Pakmanee N, Suntrarachun S, Khow O, Udomsantisuk ...
  • 31. Wu X, Feng S, Duan H, Zhang L, Zhang ...
  • 32. Wu X, Zhang L, Zhang J, Zhang C, Zhu ...
  • 33. Tebianian M, Hoseini AZ, Ebrahimi SM, Memarnejadian A, Mokarram ...
  • 34. Arend SM, Ottenhoff THM, Andersen P, Dissel JTV. Uncommon ...
  • 35. Renshaw PS, Lightbody KL, Veverka V, Muskett FW, Kelly ...
  • نمایش کامل مراجع