Detection of Cryptosporidium parvum DNA in fixed, paraffin-embedded tissue by the polymerase chain reaction.
The objective of this project was to demonstrate detection of Cryptosporidium parvum DNA in fixed, paraffin-embedded tissue using the polymerase chain reaction (PCR). DNA was purified from six samples of fixed, paraffin-embedded tissue that were histologically positive for C. parvum and used in the PCR. Previously developed oligonucleotide primers specific for C. parvum were used to amplify a 452-base target sequence, and a 20-base synthetic probe labeled with digoxigenin-11-dUTP was used to detect the amplification product by chemiluminescence. All six samples were positive by PCR; negative controls showed no amplification or hybridization. This approach could provide a sensitive and specific method for detection of parasite material in fixed, paraffin-embedded tissue samples, and prove to be of significant value in retrospective studies of archival material.
Laxer
M A
MA
Department of Infectious and Parasitic Disease Pathology, Armed Forces Institute of Pathology, Washington, DC.
D'Nicuola
M E
ME
Patel
R J
RJ
D000818:Animals; D001483:Base Sequence; D003457:Cryptosporidiosis / Q000175:diagnosis*; D016785:Cryptosporidium parvum / Q000235:genetics / Q000302:isolation & purification*; D016054:DNA, Protozoan / Q000032:analysis* / Q000737:chemistry; D005069:Evaluation Studies as Topic; D005704:Gallbladder / Q000469:parasitology; D006801:Humans; D007422:Intestines / Q000469:parasitology; D008099:Liver / Q000469:parasitology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000737:chemistry; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies
D016054:DNA, Protozoan; D015345:Oligonucleotide Probes