An oligonucleotide-ligation assay for the differentiation between Cyclospora and Eimeria spp. polymerase chain reaction amplification products.
An oligonucleotide-ligation assay (OLA) was developed and compared to a restriction fragment length polymorphism (RFLP) test for distinguishing between 294-bp polymerase chain reaction (PCR) amplification products of the 18S rRNA gene from Cyclospora and Eimeria spp. The PCR/OLA correctly distinguished between three Cyclospora, three E. tenella, and one E. mitis strains and the ratio of positive to negative spectrophotometric absorbance (A490) values for each strain ranged from 4.086 to 15.280 (median 9.5). PCR/OLA provides a rapid, reliable, spectrophotometric alternative to PCR/RFLP.
Jinneman
K C
KC
Seafood Products Research Center, Food and Drug Administration, Bothell, Washington 98041, USA. kjinnema@ora.fda.gov
Wetherington
J H
JH
Hill
W E
WE
Omiescinski
C J
CJ
Adams
A M
AM
Johnson
J M
JM
Tenge
B J
BJ
Dang
N L
NL
Wekell
M M
MM
D000818:Animals; D016054:DNA, Protozoan / Q000235:genetics; D004539:Eimeria / Q000145:classification* / Q000235:genetics; D016786:Eimeria tenella / Q000145:classification / Q000235:genetics; D016783:Eucoccidiida / Q000145:classification* / Q000235:genetics; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA
D016054:DNA, Protozoan; D015345:Oligonucleotide Probes; D012337:RNA, Ribosomal, 18S