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Nosis in 77 patients revealed that there was suppression of chronic parasitemia in 98.1 of cases in the group treated with benznidazole, in 90.4 of the group treated with nifurtimox and in 65.7 of those who received the placebo. The authors acknowledged the difficulty in assessing cure rates through this test because the individuals who used a placebo also had significant “suppression” of parasitemia [18]. Despite the questionable comparability (acute versus chronic disease) in the studied cases, we emphasize the validity of xenodiagnosis and blood culture for the short-term Title Loaded From File follow-up of patients treated during the acute phase because the frequency of positive cases obtained after 54 to 68 days of treatment are comparable with those of other authors. Cancado and Brener ?(1979) demonstrated the relative efficacy of benznidazol but found a positive xenodiagnosis after treatment in eight out of 20 chronic cases, supporting the theory of the suppressive medication effect, but not curative [19]. In cases of indeterminate Chagas disease form, treatment with benznidazol showed encouraging results from two well-controlled follow-up studies realized in Brazil and Argentina. In Brazil, the treatment with benznidazol produced negative seroconversion in children with this form of the disease. The Argentine study followed patients during four years and demonstrated a good response to benznidazol by Title Loaded From File clearance of IgG antibodies after treatment as evidence of recovery in 62 of children who used benznidazol, whereas there was no serological improvement in controls who took a placebo [20,21]. We found 47 individuals patients with negative serology, i.e., serologic cure. Significant differences were observed in the proportions of seronegative individuals acutely infected seven years ago or more and those infected less than four years. However, there were no differences between patients infected atTable 4. Parasitological and serological examinations of cases with positive PCR results.Year of acute phaseAgeQBCBlood smearXeno-diagnosisBlood cultureAcute phase titers of antibodies IgM IgG 320 160 80 0 80Follow-up phase antibodies titers1999 2000 2002 2002 200375 28 47 23 51Neg Pos Neg Neg Pos PosNeg NT Neg Neg Pos PosPos Pos Neg Pos 15857111 Pos NegNeg Pos NT Pos Pos Pos160 0 40 640 4040 40 40 40 40QBC = Quantitative buffy coat. Pos = Positive; Neg = Negative; NT = Not taken. doi:10.1371/journal.pone.0064450.tClinical Follow-Up of Acute Chagas DiseaseTable 5. PCR results and follow-up clinical status post treatment.Clinical condition Serologically cured Indeterminate or seropositive forms Cardiac form Total doi:10.1371/journal.pone.0064450.tN 47 127 5PCR-positive ( ) 0 6 (9.8) 0PCR-negative ( ) 9 (100) 55 (90.2) 2 (100)Total PCR assays 9 61 2seven years ago or more, and those infected at five or six years ago, which suggests that the period of highest negative results frequency maybe has been occurred up to the fourth year after treatment. The recommendations from endemic areas highlight that, usually, individuals undergo negative seroconversion one to five years after treatment [14]. Our results confirm these observations and also suggest that for the Amazon, this period is shorter and the retesting of patients treated should occur each six months, particularly during the fourth year after an acute infection treatment. Prognostic interpretations are difficult for 127 patients identified as seropositive without cardiac or digestive changes consistent with Chagas.Nosis in 77 patients revealed that there was suppression of chronic parasitemia in 98.1 of cases in the group treated with benznidazole, in 90.4 of the group treated with nifurtimox and in 65.7 of those who received the placebo. The authors acknowledged the difficulty in assessing cure rates through this test because the individuals who used a placebo also had significant “suppression” of parasitemia [18]. Despite the questionable comparability (acute versus chronic disease) in the studied cases, we emphasize the validity of xenodiagnosis and blood culture for the short-term follow-up of patients treated during the acute phase because the frequency of positive cases obtained after 54 to 68 days of treatment are comparable with those of other authors. Cancado and Brener ?(1979) demonstrated the relative efficacy of benznidazol but found a positive xenodiagnosis after treatment in eight out of 20 chronic cases, supporting the theory of the suppressive medication effect, but not curative [19]. In cases of indeterminate Chagas disease form, treatment with benznidazol showed encouraging results from two well-controlled follow-up studies realized in Brazil and Argentina. In Brazil, the treatment with benznidazol produced negative seroconversion in children with this form of the disease. The Argentine study followed patients during four years and demonstrated a good response to benznidazol by clearance of IgG antibodies after treatment as evidence of recovery in 62 of children who used benznidazol, whereas there was no serological improvement in controls who took a placebo [20,21]. We found 47 individuals patients with negative serology, i.e., serologic cure. Significant differences were observed in the proportions of seronegative individuals acutely infected seven years ago or more and those infected less than four years. However, there were no differences between patients infected atTable 4. Parasitological and serological examinations of cases with positive PCR results.Year of acute phaseAgeQBCBlood smearXeno-diagnosisBlood cultureAcute phase titers of antibodies IgM IgG 320 160 80 0 80Follow-up phase antibodies titers1999 2000 2002 2002 200375 28 47 23 51Neg Pos Neg Neg Pos PosNeg NT Neg Neg Pos PosPos Pos Neg Pos 15857111 Pos NegNeg Pos NT Pos Pos Pos160 0 40 640 4040 40 40 40 40QBC = Quantitative buffy coat. Pos = Positive; Neg = Negative; NT = Not taken. doi:10.1371/journal.pone.0064450.tClinical Follow-Up of Acute Chagas DiseaseTable 5. PCR results and follow-up clinical status post treatment.Clinical condition Serologically cured Indeterminate or seropositive forms Cardiac form Total doi:10.1371/journal.pone.0064450.tN 47 127 5PCR-positive ( ) 0 6 (9.8) 0PCR-negative ( ) 9 (100) 55 (90.2) 2 (100)Total PCR assays 9 61 2seven years ago or more, and those infected at five or six years ago, which suggests that the period of highest negative results frequency maybe has been occurred up to the fourth year after treatment. The recommendations from endemic areas highlight that, usually, individuals undergo negative seroconversion one to five years after treatment [14]. Our results confirm these observations and also suggest that for the Amazon, this period is shorter and the retesting of patients treated should occur each six months, particularly during the fourth year after an acute infection treatment. Prognostic interpretations are difficult for 127 patients identified as seropositive without cardiac or digestive changes consistent with Chagas.

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