کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8418118 | 1545713 | 2013 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Design and characterization of a recombinant colorimetric SAG1-alkaline phosphatase conjugate to detect specific antibody responses against Toxoplasma gondii
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
PBSIPTGNBTBCIPisopropyl-β-d-thiogalactosideSAG1Alkaline phosphatase - آلکالین فسفاتاز یا فسفاتاز قلیاییSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدT. gondii - ت.گندیدیNitro-blue tetrazolium - تترازولیم نیترو-آبیELISA - تست الیزاEnzyme-linked immunosorbent assay - تست الیزاSerodiagnosis - تشخیص سرودToxoplasma gondii - توکسوپلاسما گوندیPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریFusion protein - پروتئین فیوژن یا پروتئین همجوشی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The purpose of this study was to design a novel in vitro tool by using recombinant protein technology to detect specific antibody responses against Toxoplasma gondii in a simple, rapid and highly sensitive reagent. The surface T. gondii SAG1 protein is an important immunodominant target, which provides a great interest as a diagnostic antigen. To further exploit its immunodetection capacity, in the present study, the full length sag1 gene was inserted into the pLIP6 prokaryotic expression vector so as to produce a SAG1 antigen genetically fused to the bacterial alkaline phosphatase (AP). After expression optimization, the recombinant fusion protein folded correctly in soluble form in the periplasmic space and preserved both the AP enzymatic activity and the SAG1 immunoreactivity. Subsequently, direct-ELISA and dot-blot immunoassays were designed, using crude preparation SAG1-AP conjugate, to explore its value in serodiagnosis of human toxoplasmosis. We demonstrate that the recombinant SAG1-AP can detect specific T. gondii antibodies in one-step procedure and can successfully discriminate between T. gondii immune and non-immune patients, in agreement with the standard gold test. In conclusion, the present study shows that the genetic fusion protein provides a new tool for one-step T. gondii immunodetection, which was easily, quickly and reproducibly produced as homogeneous bi-functional reagent. Thus, this recombinant immunoconjugate is a promising marker for Toxoplasma serodiagnosis, requiring further evaluation on a larger series and could provide the basis for direct antibody capture enzyme-immunoassay for specific immunoglobulin M and G detection.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 394, Issues 1â2, 30 August 2013, Pages 107-114
Journal: Journal of Immunological Methods - Volume 394, Issues 1â2, 30 August 2013, Pages 107-114
نویسندگان
Nouha Chahed Bel-Ochi, Aïda Bouratbine, Mohamed Mousli,