Bindu Lakshmanan1, Jain Jose2, Amrutha Anand2, M N Priya2. 1. Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Kerala Veterinary and Animal Sciences University, Mannuthy, Wayanad, Kerala, 680651, India. bindul@kvasu.ac.in. 2. Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Kerala Veterinary and Animal Sciences University, Mannuthy, Wayanad, Kerala, 680651, India.
Abstract
PURPOSE: Visceral schistosomosis is an economically important trematode infection caused by Schistosoma spindale and S. indicum in among ruminants. The lack of sensitive diagnostic tools has often led to underestimation of the prevalence in live animals. A sensitive copro-PCR targeting partial mitochondrial gene was developed to detect Schistosoma spp. However, this protocol could not differentiate between the two species. This study was conducted to explore the possibility of species differentiation using restriction fragment length polymorphism of PCR products (PCR- RFLP). METHODS: Polymerase chain reaction was carried out to amplify mitochondrial gene of adult S. spindale and S. indicum. Copro PCR was done with schistosome-positive faecal samples. A novel PCR-RFLP was designed targeting the Hpy166II recognition sequence in the mitochondrial gene sequence of S. indicum. RESULT: The PCR using primers targeting the mitochondrial gene of S. spindale and S. indicum amplified a distinct product of approximately 454 bp with adult fluke as well as faecal DNA, which upon RFLP with Hpy166II yielded 330 bp and 124 bp products with S. indicum amplicons alone. CONCLUSION: The novel PCR-RFLP possesses the potential to be used in epidemiological surveys among bovines and in snail intermediate hosts to screen for S. spindale and S. indicum infection.
PURPOSE: Visceral schistosomosis is an economically important trematode infection caused by Schistosoma spindale and S. indicum in among ruminants. The lack of sensitive diagnostic tools has often led to underestimation of the prevalence in live animals. A sensitive copro-PCR targeting partial mitochondrial gene was developed to detect Schistosoma spp. However, this protocol could not differentiate between the two species. This study was conducted to explore the possibility of species differentiation using restriction fragment length polymorphism of PCR products (PCR- RFLP). METHODS: Polymerase chain reaction was carried out to amplify mitochondrial gene of adult S. spindale and S. indicum. Copro PCR was done with schistosome-positive faecal samples. A novel PCR-RFLP was designed targeting the Hpy166II recognition sequence in the mitochondrial gene sequence of S. indicum. RESULT: The PCR using primers targeting the mitochondrial gene of S. spindale and S. indicum amplified a distinct product of approximately 454 bp with adult fluke as well as faecal DNA, which upon RFLP with Hpy166II yielded 330 bp and 124 bp products with S. indicum amplicons alone. CONCLUSION: The novel PCR-RFLP possesses the potential to be used in epidemiological surveys among bovines and in snail intermediate hosts to screen for S. spindale and S. indicum infection.
Authors: O P Akinwale; T Laurent; P Mertens; T Leclipteux; D Rollinson; R Kane; A Emery; M B Ajayi; D O Akande; T W Fesobi Journal: Mol Biochem Parasitol Date: 2008-04-12 Impact factor: 1.759
Authors: N Sandoval; M Siles-Lucas; J L Pérez-Arellano; C Carranza; S Puente; J López-Abán; A Muro Journal: Parasitology Date: 2006-07-12 Impact factor: 3.234