Detection of Paragonimus heterotremus eggs in experimentally infected cats by a polymerase chain reaction-based method.
A polymerase chain reaction (PCR) procedure for the detection of Paragonimus heterotremus eggs in stool samples was developed and compared with Stoll's egg count method. The primers were designed on the basis of a previously constructed pPH-13-specific DNA probe, which produced an approximate 0.5-kb amplified product. This PCR method could detect as few as 5 eggs in 0.6 g of artificially inoculated feces of a healthy control cat or as little as 1 x 10(-4) ng of P. heterotremus genomic DNA. The assay had 100% sensitivity in all infected cats. The method did not yield an approximate 0.5-kb product with DNA from other parasites such as Gnathostoma spinigerum, Trichinella spiralis, Fasciola gigantica, Echinostoma malayanum, Opisthorchis viverrini, Dirofilaria immitis, and Taenia saginata; exceptions were Paragonimus siamensis and Paragonimus westermani. In addition, no genomic DNA from Escherichia coli, Burkholderia pseudomallei, Acinetobacter anitratus, Mycobacterium tuberculosis, Staphylococcus aureus, beta-Streptococcus grA, and Proteus mirabilis or from the vertebrate and invertebrate hosts of P. heterotremus was amplified in the PCR assay. This assay has great potential for application in clinical epidemiological studies.
Intapan
Pewpan M
PM
Departmentsof Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. pewpan@kku.ac.th
Wongkham
Chaisiri
C
Imtawil
Kanokwan J
KJ
Pumidonming
Wilawan
W
Prasongdee
Thidarat K
TK
Miwa
Masanao
M
Maleewong
Wanchai
W
D000818:Animals; D001483:Base Sequence; D002371:Cat Diseases / Q000175:diagnosis / Q000469:parasitology*; D002415:Cats; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes; D018509:DNA, Helminth / Q000737:chemistry / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology*; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D010237:Paragonimiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D010238:Paragonimus / Q000235:genetics / Q000302:isolation & purification*; D010270:Parasite Egg Count / Q000379:methods / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000662:veterinary
D017931:DNA Primers; D015342:DNA Probes; D018509:DNA, Helminth