Direct Identification of Trypanosoma Cruzi Natural Clones in Vectors and Mammalian Hosts by Polymerase Chain Reaction Amplification

Selection of Trypanosoma cruzi clonal genotypes (clonet 20 and 39) isolated from Bolivian triatomines following subculture in liquid medium

“…After electrophoresis the PCR products were transferred on to membranes and successively hybridized with the specific clonet probes 20 and 39 (Veas et al 1991, Brenière et al 1992. Total DNAs were purified by phenol/chloroform extraction from parasite pellets of each stock and similarly, the clonets were detected by hybridization of the PCR products amplified from total DNAs.…”

“…Identification of clonet 20 and 39 by PCR/ hybridization -The procedures for feces collection, extraction and PCR were according to Brenière et al (1992Brenière et al ( , 1995, using primers (Genset, Paris, France) chosen to amplify the hyper variable region of the kinetoplast minicircles (HVRm). After electrophoresis the PCR products were transferred on to membranes and successively hybridized with the specific clonet probes 20 and 39 (Veas et al 1991, Brenière et al 1992.…”

Comparison of Polymerase Chain Reaction Methods for Reliable and Easy Detection of Congenital Trypanosoma Cruzi Infection

“…16 However, such PCRs require partial disruption of the dense kinetoplast minicircle with either a nuclease 11 or by boiling the sample in the presence of strong chaotropic agents such as guanidine chloride 17 to release linear and amplifiable kDNA fragments. In addition, the quantitative reproducibility of such treatments in releasing kDNA minicircles of parasites was not assessed.…”

“…18 The first T. cruzi PCRs were able to detect only large quantities of DNA and required extraction of large volumes of blood and a supplementary hybridization step or the use of a hot start PCR to increase their sensitivities and specificities. 5,13,14,16,[19][20][21] Moreover, the diagnostic application of a PCR also requires the detection of all T. cruzi strains that can infect patients. Recently, T. cruzi was divided in two major lineages.…”

DNA evidence of Trypanosoma cruzi in the Chilean wild vector Mepraia spinolai (Hemiptera: Reduviidae)

“…Detection of T. cruzi can be carried out through different methodologies such as direct microscopic observation, hemoculture, xenodiagnosis, and in the last decade the polymerase chain reaction (PCR). It is well known that PCR-based detection from feces or urine of reduviid bugs, and blood samples from mammals is more efficient than the other techniques (Moser et al 1989, Breniere et al 1992, Russomando et al 1992. However, scarce information has been reported about infection levels of wild triatomine populations using molecular techniques.…”