کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9443002 | 1302616 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Neospora caninum: Quantification of DNA in the blood of naturally infected aborted and pregnant cows using real-time PCR
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کلمات کلیدی
Tris–borate–EDTArRNATNFUngNeoRIgGT helperBVDVTBEMgCl2IgEDMSO - DMSODNA - DNA یا اسید دزوکسی ریبونوکلئیکReal-time PCR - PCR به موقعenzyme linked immunosorbent assay - آنزیم تست ایمونوسیورسانس مرتبط استBasic Local Alignment Search Tool - ابزار جستجوی محلی سازگاری محلیdeoxyribonucleic acid - اسید deoxyribonucleicribosomal ribonucleic acid - اسید ریبونوکلئیک ریبوزومیUltraviolet - اشعه فرابنفشBlast - انفجارImmunoglobulin E - ایمونوگلوبولین Eimmunoglobulin G - ایمونوگلوبولین Ginterferon - اینترفرونIFN - اینترفرون هاinterleukin - اینترلوکینpregnant - باردارT. gondii - ت.گندیدیELISA - تست الیزاToxoplasma gondii - توکسوپلاسما گوندیbase pairs - جفت پایهDimethyl sulfoxide - دیمتیل سولفواکسیدAbortion - سقط جنینtumour necrosis factor - عامل نکروز تومورNeospora caninum - نئوسپورا نئوسپوراpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازBovine viral diarrhoea virus - ویروس اسهال ویروسی گاوParasitaemia - پارازیتمیسمMagnesium chloride - کلرید منیزیم
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
انگل شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
This study quantified Neospora caninum DNA in the blood and brain of pregnant and aborted heifers by monitoring PCR product formation in real-time using SYBR Green I, a double-stranded DNA-binding dye. Primers were designed to amplify a 188Â bp product specific to N. caninum from the Nc-5 gene fragment of N. caninum. Similarly, a 71Â bp product was amplified from the 28S rRNA gene of bovine genomic DNA that served as a control. Agarose gel electrophoresis and analysis of the melting curve for PCR products showed that both primer pairs were specific to their targets. Standard curves were generated for both bovine and N. caninum genomic DNA, and were used to compute the relative concentration of parasite to bovine DNA in the test samples. The concentration of N. caninum DNA in 1Â ng of bovine genomic DNA obtained from blood ranged between 0.097Â ng at the 1st week of the observation and 0Â ng at the 15th week in aborted cows. In pregnant cows, the values ranged between 0.080Â ng at the 1st week and 0.155Â ng at the 15th week of observation. There was a sustained decrease of DNA concentration in the aborted group after abortion and an increase in DNA concentration in the pregnant group. Comparison of parasite DNA in blood and brain of infected heifers showed a higher DNA concentration in brain than in blood. This study shows that N. caninum DNA can be quantified to obtain the relative concentration of parasite DNA to host genomic DNA in blood. This technique allows testing of a large number of samples at one time and can be done without the need for slaughter of tested animals.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Experimental Parasitology - Volume 110, Issue 1, May 2005, Pages 48-55
Journal: Experimental Parasitology - Volume 110, Issue 1, May 2005, Pages 48-55
نویسندگان
C.M. Okeoma, K.M. Stowell, N.B. Williamson, W.E. Pomroy,