development and evaluation of a sensitive pcr-elisa system for detection of schistosoma infection in feces开发和评估一个敏感pcr-elisa系统检测血吸虫感染的粪便.pdfVIP

Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces ´ 1 ´ 1 2 ´ Luciana Inacia Gomes , Letıcia Helena dos Santos Marques , Martin Johannes Enk , Maria Claudia de Oliveira1, Paulo Marcos Zech Coelho2, Ana Rabello 1* ´ ´ ´ ˜ ´ 1 Laboratorio de Pesquisas Clınicas, Centro de Pesquisas Rene Rachou, Fundac¸ao Oswaldo Cruz (Fiocruz), Belo Horizonte, Minas Gerais, Brazil, 2 Laboratorio de ´ ˜ Esquistossomose, Centro de Pesquisas Rene Rachou, Fundac¸ao Oswaldo Cruz (Fiocruz), Belo Horizonte, Minas Gerais, Brazil Abstract Background: A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed to overcome the need for sensitive techniques for the efficient diagnosis of Schistosoma infection in endemic settings with low parasitic burden. Methodology/Principal Findings: This system amplifies a 121-base pair tandem repeat DNA sequence, immobilizes the resultant 59 biotinylated product on streptavidin-coated strip-well microplates and uses anti-fluorescein antibodies conjugated to horseradish peroxidase to detect the hybridized fluorescein-labeled oligonucleotide probe. The detection limit of the Schistosoma PCR-ELISA system was determined to be 1.3 fg of S. mansoni genomic DNA (less than the amount found in a single cell) and estimated to be 0.15 S. mansoni eggs per gram of feces (fractions