Multiplex PCR for both Identification of Brazilian Biomphalaria Species (Gastropoda: Planorbidae) and Diagnosis of Infection by Schistosoma mansoni (Trematoda: Schistosomatidae)

• Published in: The Journal of parasitology Vol. 92; no. 2; pp. 401 - 403
• Main Authors: Jannotti-Passos, Liana K, Magalhães, Kelly Grace, Carvalho, Omar S, Vidigal, Teofânia H. D A
• Format: Journal Article
• Language: English
• Published: Lawrence, KS American Society of Parasitologists 01.04.2006; Allen Press Inc
• Subjects: Animals; Annealing; Biological and medical sciences; Biological taxonomies; Biomphalaria; Biomphalaria - classification; Biomphalaria - genetics; Biomphalaria - parasitology; Brazil; Cercariae; Diagnosis; DNA Primers - chemistry; DNA, Mitochondrial - analysis; DNA, Ribosomal Spacer - analysis; Fundamental and applied biological sciences. Psychology; Gastropoda; Gels; General aspects; General aspects and techniques. Study of several systematic groups. Models; Identification; Infections; Invertebrates; Mitochondrial DNA; Mollusca; Mollusks; Morphology; Multiplexing; Parasitology; Planorbidae; Polyacrylamide; Polymerase chain reaction; Polymerase Chain Reaction - methods; Reproducibility of Results; Research Notes; Ribosomal DNA; RNA, Ribosomal, 18S - genetics; Schistosoma mansoni; Schistosoma mansoni - genetics; Schistosoma mansoni - isolation & purification; Schistosomatidae; Schistosomiasis; Sensitivity and Specificity; Snails; Species; Species Specificity; Trematoda; Brazil; Schistosoma mansoni; Plathelmintha; Diagnosis (taxonomy); Parasite; Gastropoda; Trematoda; Infection; Polymerase chain reaction; Pulmonata; Biomphalaria; Helmintha; Intermediate host; Invertebrata; Mollusca
• ISSN: 0022-3395; 1937-2345
• DOI: 10.1645/GE-593R1.1

A simple and single-step technique based on multiplex PCR (multiplex polymerase chain reaction) has been developed for simultaneous identification of Brazilian Biomphalaria species, the intermediate hosts of Schistosoma mansoni, and their diagnosis of infection by the trematode. We used species-specific primers directed both to the internal transcribed spacer 2 (ITS2) of ribosomal DNA from 3 of the S. mansoni host species and to the mitochondrial DNA (mtDNA) from the trematode. Those primers were used simultaneously in a single multiplex-PCR reaction, and template DNA was obtained from S. mansoni– infected and noninfected snails. The results were visualized in silver stained polyacrylamide gels, revealing the presence of specific bands. The methodology has shown to be efficient, fast, and reproducible for Biomphalaria species identification and diagnosis of snails infected by S. mansoni during prepatent periods.