Chronic Chagas disease: PCR-xenodiagnosis without previous microscopic observation is a useful tool to detect viable Trypanosoma cruzi

We evaluate the elimination of the microscopic stage of conventional xenodiagnosis (XD) to optimize the parasitological diagnosis of Trypanosoma cruzi in chronic Chagas disease. To this purpose we applied under informed consent two XD cages to 150 Chilean chronic chagasic patients. The fecal samples (FS) of the triatomines at 30, 60 and 90 days post feeding were divided into two parts: in one a microscopic search for mobile trypomastigote and/or epimastigote forms was performed. In the other part, DNA extraction-purification for PCR directed to the conserved region of kDNA minicircles of trypanosomes (PCR-XD), without previous microscopic observation was done. An XD was considered positive when at least one mobile T. cruzi parasite in any one of three periods of incubation was observed, whereas PCR-XD was considered positive when the 330 bp band specific for T. cruzi was detected. 25 of 26 cases with positive conventional XD were PCR-XD positive (concordance 96.2%), whereas 85 of 124 cases with negative conventional XD were positive by PCR-XD (68.5%). Human chromosome 12 detected by Real-time PCR used as exogenous internal control of PCR-XD reaction allowed to discounting of PCR inhibition and false negative in 40 cases with negative PCR-XD. Conclusion: PCR-XD performed without previous microscopic observation is a useful tool for detection of viable parasites with higher efficiency then conventional XD.

Enfermedad de Chagas crónica: ausencia de Triatoma infestans intradomiciliario y persistencia de Trypanosoma cruzi circulante post-terapia / Chronic Chagas disease: absence of intradomiciliary Triatomatoma infestans and persistence of circulating post-therapy Trypanosoma cruzi

En el año 1999, se certificó en Chile la interrupción de la transmisión vectorial de la enfermedad de Chagas. Por otra parte, el desarrollo de políticas habitacionales, que incluyen zonas rurales endémicas, han logrado importantes avances en grupos humanos que presentaban graves deficiencias de habitabilidad. En esta situación epidemiológica, se estudió 10 años después de la terapia, a 37 chagásicos crónicos procedentes de zonas rurales de la IV Región, mediante encuesta epidemiológica (pre y post-terapia) y evaluación parasitológica (post-terapia). La encuesta incluyó: conocimiento del vector, triatominos en el domicilio actual, antecedentes de haber sido picado por triatominos y material de la construcción de la vivienda. La evaluación parasitológica se realizó mediante xenodiagnóstico (XD) (37 casos) y Reacción en Cadena de la Polimerasa (PCR) en sangre periférica (34 casos). Los resultados de la encuesta epidemiológica evidenciaron cambios importantes en la calidad de la vivienda rural de los chagásicos tratados, mientras que, en relación a la parasitemia, se detectó Trypanosoma cruzi circulante en el 89,2 por ciento de los casos (37,8 por ciento y 88,2 por ciento, mediante XD y PCR, respectivamente). Se concluye que la persistencia del parásito en condiciones post-terapia, no está relacionada con la presencia de Triatoma infestans intradomiciliario.

Rendimiento del xenodiagnóstico, según el número de cajas utilizadas en 1.181 personas con infección chagásica crónica diagnosticada mediante la reacción de hemaglutinación indirecta / Yielding of xenodiagnosis, according to the number of boxes utilized, in 1.181 persons with chronic chagasic infection diagnosed by means of indirect hemagglutination reaction

A study for evaluation of the yielding of xenodiagnosis (XD) in 1,181 persons with a previous positive indierect hemagglutination test (IHAT) for Trypanosoma cruzi infection was carried out. The infections of these people was detected during epidemiological surveys performed in rural-periurban and urban sections of the endemo-enzootic area of Chagas' disease in Chile, which involves the first seven, out of the thirteen political-administrative regions of the country. The sex distribution was 75.0% females and 25.0% males, varying the ages between 2 and 80 years. According to individual and geographical possibilities each person was submitted to 1-8 XD, which consisted in cylindrical wooden boxes containing seven third instar nymphs, laboratory reared, of Triatoma infestans. The boxes, covered with a pieces of tulle fixed with a rubber band were applied on the skin of the posterior side of the arm of the subject to be examined, holdea with a linen bracelet during 25-30 min. After the insects were fed the boxes were maintained in the laboratory at 27-C and 85% relative environmental humidity. Posteriorly, all the nymphs of each box were examined at 30, 60 and 90 days after application. A drop of abdominal content of each of them, homogenized with a drop of saline, was examined at the microscope looking for T. cruzi. XD resulted positive in 503 (42.6%) people. The positiveness of XD showed a trend of increasing according to the number of boxes used, from 11.4% with one to 51.6% with six. Under a practical point of view, the simultaneous application of four XD boxes seems to be advisable. In relation to the IHAT for Chagas' infection, the percentages of XD positivity increased from 39.8% in those people with lower titers (16-128) to 61.7% in those with higher titers (256-1,000)