Detection of Cryptosporidium parvum and C. muris Oocysts in Spiked Backwash Water using Three PCR-Based Protocols

Detection of Infectious Cryptosporidium parvum Oocysts in Surface and Filter Backwash Water Samples by Immunomagnetic Separation and Integrated Cell Culture-PCR

“…The current U.S. Environmental Protection Agency Information Collection Rule method (23) and proposed method 1622 (21) for the detection of Cryptosporidium recovered from water samples do not specifically detect the human pathogen Cryptosporidium parvum or determine the viability or infectivity of recovered oocysts. Recently, several PCR-based methods for the detection of C. parvum have been described (4,6,7,9,20,24); these include an infectivity assay based on in vitro cell culturing of the parasite with Caco-2 cells and detection of C. parvum-infected cells by targeting C. parvum hsp70 mRNA by use of reverse transcription (RT)-PCR (18). We have developed another strategy for a C. parvum infectivity assay which integrates immunomagnetic separation (IMS) to recover Cryptosporidium oocysts from water samples with in vitro cell culturing with human ileocecal adenocarcinoma HCT-8 cells and detection of C. parvum-infected cells by targeting C. parvum hsp70 DNA by use of standard PCR (CC-PCR) (5).…”

Padronização de uma reação em cadeia pela polimerase em nested (nested-PCR) para detecção e diferenciação das espécies de Cryptosporidium spp e caracterização molecular de Cryptosporidium isolados de roedores sinantrópicos

“…Cryptosporidium é um gênero de protozoário cosmopolita pertencente ao filo (BLACK et al, 1996;BONNIN;FOURMAUX;DUBREMETZ, 1996;SPANO et al, 1997;GIBBONS;RIGI;AWAD-EL-KARIEM, 1998;XIAO et al, 2002 Baseado em FAYER et al, 1997FAYER et al, , 2000FAYER et al, , 2001FAYER et al, , 2004FAYER et al, , 2006, Monis e Thompson (2003), Morgan et al, (1999), Xiao et al, (2000). (A) Abomaso, (ID) Intestino Delgado, (E) Estômago, (IG) Intestino grosso, (B) Bursa de Fabrícius, (C MORGAN et al, 2000;XIAO et al, 2000;FAYER et al, 2001) • Peneirou-se 50 g de fezes em tamises de 0,5 mm, 150 um e 45 um, sendo diluídas ao poucos com 350 mL de água destilada.…”