DETECTION OF TRYPANOSOMA-CRUZI IN BLOOD SPECIMENS OF CHRONIC CHAGASICPATIENTS BY POLYMERASE CHAIN-REACTION AMPLIFICATION OF KINETOPLAST MINICIRCLE DNA - COMPARISON WITH SEROLOGY AND XENODIAGNOSIS
Ha. Avila et al., DETECTION OF TRYPANOSOMA-CRUZI IN BLOOD SPECIMENS OF CHRONIC CHAGASICPATIENTS BY POLYMERASE CHAIN-REACTION AMPLIFICATION OF KINETOPLAST MINICIRCLE DNA - COMPARISON WITH SEROLOGY AND XENODIAGNOSIS, Journal of clinical microbiology, 31(9), 1993, pp. 2421-2426
A panel of 114 blood samples from chronic chagasic patients and noncha
gasic patients was screened for Trypanosoma cruzi by xenodiagnostic, s
erologic, and polymerase chain reaction (PCR) amplification tests. Blo
od samples were preserved in a guanidine-EDTA buffer, and total blood
DNA was isolated after chemical nuclease cleavage with 1,10-phenanthro
line-copper ion and used as a template for PCR amplification of the co
nserved and variable regions of T. cruzi minicircle molecules. The PCR
products were screened by Southern blot hybridization with a digoxige
nin-labeled oligonucleotide probe specific for the conserved region of
the minicircle. The method showed a sensitivity of 100% compared witb
the serologic test. In addition, all of the serology-positive, xenodi
agnosis-negative samples were positive by PCR. This demonstrates that
PCR amplification of T. cruzi kinetoplast minicircle DNA could replace
xenodiagnosis for evaluation of parasitemia in chronic chagasic patie
nts and could serve as a complement for serologic testing in the scree
ning of blood bank donors.