TY  - JOUR
T1  - Detection and Differentiation of <I>Echinococcus granulosus-</I>Complex Using A Simple PCR-Based Assay
AU - , Ahmed M.A. Osman AU - , Imadeldin E. Aradaib AU - , Abdel-Latif K. Ashmaig AU - , Ahmed A. Gameel 
JO  - International Journal of Tropical Medicine
VL  - 4
IS  - 1
SP  - 21
EP  - 26
PY  - 2009
DA  - 2001/08/19
SN  - 1816-3319
DO  - ijtmed.2009.21.26
UR  - https://makhillpublications.co/view-article.php?doi=ijtmed.2009.21.26
KW  - Hydatid disease
KW  -cystic echinococcosis
KW  -Echinococcus granulosus
KW  -complex
KW  -genotyping
KW  -molecular diagnosis
KW  -PCR
AB  - A nested Polymerase Chain Reaction (PCR) assay, for detection of intact and calcified hydatid cysts of <I>Echinococcus granulosus</I> (EG)-complex, was developed and evaluated. The NADH dehydrogenase 1 gene was used as a target DNA for PCR amplification. Two pairs of primers, (EGL 1 and EGR 2) and (EGL 3 and EGR 4), were designed and used in 2 amplification steps. First, the outer pair of primers (EGL 1 and EGR 2), derived from a highly conserve region of NADH 1 gene produced a primary 435 base pair (bp) PCR product from different stains of EG-complex including, sheep strain (genotype) designated (G 1), Cattle strain (G 5), camel strain (G 6) and pig strain (G 7). Second, a pair of internal (nested) primers (EGL 3 and EGR 4), designed internal to the annealing sites of primers (EGL 1 and EGR 2), produced a 276 bp PCR product. The primary 435 bp and the nested 276 bp EG specific PCR products were easily identified following visualization onto an ethidium bromide-stained agarose gel. However, the primary or the nested PCR products were not amplified from DNA extracted from <I>Cysticercus tenuicollis</I> and <I>Coenurus cerebralis</I>, the larval stages of the adult dog cestodes <I>Taenia hydatigena</I> and <I>T. multiceps</I>, respectively. The described nested PCR assay could be used to detect sheep, cattle and camel strains of EG-complex circulating in Sudan. This PCR assay could also, be used for rapid detection and differentiation of EG-complex from other related cestodes. This assay should be considered during an epidemiological survey of the disease in areas of endemicity.
ER  -