Comparative efficacy of conventional primer sets in detection of Cryptosporidium parvum for diagnostic use.
In this study, the sensitivity and specificity of different previously described primer sets for Cryptosporidium parvum detection by polymerase chain reaction (PCR) was evaluated. For this purpose, the primer sets defined by Cacciò et al. (FEMS Microbiol Lett 170(1):173-179, 1999) (tub), Widmer et al. (Appl Environ Microbiol 64(11):4477-4481, 1998) (btub) and Rochelle et al. (Appl Environ Microbiol 63:2029-2037, 1997) (cphsp), respectively, were used. Deoxyribonucleic acid (DNA) was isolated from three different sample materials: (1) from the faeces of an experimentally C. parvum-infected calf, (2) from purified C. parvum oocysts, and (3) from C. parvum-infected HCT-8 cell cultures. The DNA samples were subjected to PCR reactions with each of the three given primer sets to investigate sensitivity and suitability for routine use. The primers described by Cacciò et al. (FEMS Microbiol Lett 170(1):173-179, 1999) (TUB) were superior regarding sensitivity and specificity in terms of detection of C. parvum in faeces, in purified oocysts and also in cell culture, and may thus be applied for routine diagnostic use in common sample materials.
Kar
Sirri
S
Department of Parasitology, Faculty of Veterinary Medicine, University of Ankara, 06110 Diskapi, Ankara, Turkey.
Daugschies
Arwid
A
Bangoura
Berit
B
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology; D002460:Cell Line; D003457:Cryptosporidiosis / Q000175:diagnosis* / Q000469:parasitology; D016785:Cryptosporidium parvum / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics*; D005243:Feces / Q000469:parasitology; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers