Polymerase Chain Reaction for Chronic Trypanosoma cruzi Infection Yields Higher Sensitivity in Blood Clot Than Buffy Coat or Whole Blood Specimens

Fitzwater, Sean; Calderón, Maritza; LaFuente, Carlos; Galdos-Cardenas, Gerson; Ferrufino, Lisbeth; Verástegui, Manuela; Gilman, Robert H.; Bern, Caryn; Bolivia,

doi:10.4269/ajtmh.2008.79.768

Cited by 42 publications

(41 citation statements)

References 15 publications

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“…The use of clot was based on an analysis that demonstrated higher sensitivity for clot compared to buffy coat or whole blood mixed with guanidine [30]. DNA was extracted following a standard phenol-chloroform protocol [31].…”

Section: Methodsmentioning

confidence: 99%

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CongenitalTrypanosoma cruziTransmission in Santa Cruz, Bolivia

et al. 2009

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Background We conducted a study of congenital Trypanosoma cruzi infection in Santa Cruz, Bolivia. Our objective was to apply new tools to identify weak points in current screening algorithms, and find ways to improve them. Methods Women presenting for delivery were screened by rapid and conventional serological tests. For infants of infected mothers, blood specimens obtained on days 0, 7, 21, 30, 90, 180, and 270 were concentrated and examined microscopically; serological tests were performed for the day 90, 180, and 270 specimens. Maternal and infant specimens, including umbilical tissue, were tested by polymerase chain reaction (PCR) targeting the kinetoplast minicircle and by quantitative PCR. Results Of 530 women, 154 (29%) were seropositive. Ten infants had congenital T. cruzi infection. Only 4 infants had positive results of microscopy evaluation in the first month, and none had positive cord blood microscopy results. PCR results were positive for 6 (67%) of 9 cord blood and 7 (87.5%) of 8 umbilical tissue specimens. PCR-positive women were more likely to transmit T. cruzi than were seropositive women with negative PCR results (P < .05). Parasite loads determined by quantitative PCR were higher for mothers of infected infants than for seropositive mothers of uninfected infants (P < .01). Despite intensive efforts, only 58% of at-risk infants had a month 9 specimen collected. Conclusions On the basis of the low sensitivity of microscopy in cord blood and high rate of loss to follow-up, we estimate that current screening programs miss one-half of all infected infants. Molecular techniques may improve early detection.

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Section: Methodsmentioning

confidence: 99%

“…DNA was extracted following a standard phenol-chloroform protocol [31]. Amplifications were performed using the 121/122 kinetoplast DNA primer set, as described elsewhere [30, 32]. …”

Section: Methodsmentioning

confidence: 99%

CongenitalTrypanosoma cruziTransmission in Santa Cruz, Bolivia

et al. 2009

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“…The extracted DNA was resuspended in 100 L of 10 mmol/L Tris-HCl, 1 mmol/L EDTA. 39 A PCR targeting the kinetoplast DNA of T. cruzi was performed as previously described using primers 121 (5=-AAATAATGTACGGGKGAGATGCATGA-3=) and 122 (5=-GGTTCGATTGGGGTTGGTGTAATATA-3=). 40,41 We included internal control primers that amplify a 71-bp fragment.…”

Section: Dna Extraction and Pcrmentioning

confidence: 99%

Cavia porcellus as a Model for Experimental Infection by Trypanosoma cruzi

Castro-Sesquen

Gilman

Yauri

et al. 2011

The American Journal of Pathology

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The guinea pig (Cavia porcellus) is a natural reservoir for Trypanosoma cruzi but has seldom been used as an experimental infection model. We developed a guinea pig infection model for acute and chronic Chagas disease. Seventy-two guinea pigs were inoculated intradermally with 10 4 trypomastigotes of T. cruzi strain Y (experimental group); 18 guinea pigs were used as control group. Eight animals from the experimental group and two from the control group were Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is one of the most important parasitic infections of the Americas. Despite successful vector control programs in many countries, the disease remains a major public health problem in Latin America, with 8 to 10 million people currently infected, an annual incidence of 65,000 new cases in 15 countries, and 14,000 deaths associated with the infection per year. 1 Transmission to the mammalian host occurs when a T. cruzi-infected triatomine vector deposits infectious trypomastigotes in feces during a blood meal and the parasites invade through the bite wound or intact mucosal membranes. 2 Trypomastigotes can infect virtually any nucleated cell, where they transform to amastigotes and replicate in the host cell cytoplasm. Amastigotes convert back to trypomastigotes inside the cell and then rupture the cell and circulate in the bloodstream. After weeks, the host immune response usually controls the acute infection but does not completely clear the parasite. This results in lifelong chronic infection of the host.The initial weeks after T. cruzi infection are characterized by a high-level parasitemia and symptoms that vary from mild nonspecific febrile illness to life-threatening myocarditis and/or meningoencephalitis. 2 Acute symptoms and detectable parasitemia resolve spontaneously within 2 to 3 months, and the infected individual passes into the chronic phase of the disease. Nearly all individuals in the early period of chronic infection are asymptomatic and are said to have the indeterminate form of the disease. In most individuals, the infection remains silent for life and is detectable only by anti-T. cruzi serologic tests and, in a proportion of individuals, by molecular Supported by NIH training grant in infectious and tropical diseases 5 T35 AI065385 and NIH grant 1R01AI087776-01.

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“…Ease-of-use serological screening tests have been developed [230]. Molecular techniques have been standardized [231], and have significantly improved the early diagnosis of congenital Chagas infection [232,233]. Progress in molecular techniques has allowed confirmation of persistent T. cruzi infection in the tissues of patients chronically infected and have underlined the need for parasite elimination at any moment of disease evolution [234,235].…”

Section: Developments Since 2000mentioning

confidence: 99%

Therapy of vector-borne protozoan infections in nonendemic settings

Bottieau

Vekemans

Gompel

2011

Expert Review of Anti-infective Therapy

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Vector-borne protozoan infections are responsible for a wide variety of illnesses (mainly malaria, trypanosomiasis and leishmaniasis) affecting tropical and subtropical areas, but increasingly diagnosed in nonendemic settings. This article summarizes the therapeutic developments for these conditions during the past decade and focuses specifically on treatment recommendations for returning travelers and migrants. The treatment of malaria has known the most spectacular improvements. Progress in the management of leishmaniasis and trypanosomiasis has also been substantial and includes introduction of new drugs into clinical practice, combinations of existing drugs, or new laboratory tools for treatment monitoring as well as extension of treatment indications to new groups of patients. Serious gaps still exist in terms of effectiveness and tolerance. Since the research pipeline is very limited for the coming 5-10 years, optimized combinations of existing drugs need to be urgently explored.

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Polymerase Chain Reaction for Chronic Trypanosoma cruzi Infection Yields Higher Sensitivity in Blood Clot Than Buffy Coat or Whole Blood Specimens

Cited by 42 publications

References 15 publications

CongenitalTrypanosoma cruziTransmission in Santa Cruz, Bolivia

CongenitalTrypanosoma cruziTransmission in Santa Cruz, Bolivia

Cavia porcellus as a Model for Experimental Infection by Trypanosoma cruzi

Therapy of vector-borne protozoan infections in nonendemic settings

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