Direct isolation of DNA from patient stools for polymerase chain reaction detection of Cryptosporidium parvum.
Although polymerase chain reaction (PCR) can sensitively detect parasitic or other infections, its use with fecal samples is extremely limited, primarily because of the presence of substances that inhibit DNA extension. Here an improved protocol is reported for directly isolating DNA from aged or fresh formalin-fixed stools, which can then be used to detect Cryptosporidium parvum by nested PCR. This method is highly reproducible, sensitive, and specific. It detects <1 pg of C. parvum DNA in human stool, and there are no cross-reactions with other parasites commonly found there.
Zhu
G
G
Wadsworth Center, New York State Department of Health, Albany 12208, USA.
Marchewka
M J
MJ
Ennis
J G
JG
Keithly
J S
JS
D000818:Animals; D003457:Cryptosporidiosis / Q000175:diagnosis*; D016785:Cryptosporidium parvum / Q000302:isolation & purification; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification*; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000469:parasitology; D006801:Humans; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D016054:DNA, Protozoan