Abstract

Chagas disease, a neglected protozoal disease endemic in Latin America, is also currently considered an emerging threat in nonendemic areas because of population movements. The detection of Trypanosoma cruzi DNA is increasingly being considered as important evidence to support Chagas disease diagnoses. However, further performance evaluation of molecular assays is useful for a standardization of strategy considering the whole process in routine diagnosis, especially for the different settings such as endemic and nonendemic countries. Seventy-five samples were collected from subjects screened for Chagas disease in Italy. The DNA was isolated from blood using automated extraction. We evaluated the performance of the commercial RealCycler® CHAG kit (pmPCR) based on satellite DNA (SatDNA) and of an in-house real-time PCR (ihPCR) targeting Sat and kinetoplast (k) DNAs, using the concordance of two serology assays as a reference standard. The sensitivity of kDNA and SatDNA tests by ihPCR and SatDNA by pmPCR were 14.29% (95% confidence interval (CI) 6.38 to 26.22), 7.14% (95% CI 1.98 to 17.29), and 7.14% (95% CI 1.98 to 17.29), respectively. Specificity was 100% for all PCR assays and targets. Overall, our results suggest that the preferred approach for clinical laboratories is to combine the kDNA and SatDNA as targets in order to minimize false-negative results increasing sensitivity.

Highlights

  • Chagas disease had been described for the first time in 1909 by the Brazilian medical doctor Carlos Chagas

  • Tnhoene1n9dsepmeiccimcoeunnstrfyr.om patients diagnosed as negative by our routine diagnosis were confirmed (atdaa(1dconscis7aiaaaan1.lgn2lclg7ecyn9uen.guTz2)goloa,lea9hasatsdt)irteities,ievesv,db1rsd,ewe9eypwbwsbewsabpaaacpsiosseittseteih7tech7vchd.td1i.bei1mbPv4looo4yo%eCnetn%.tlhnRhycc(T.s(P9llPt9hiif5CeTnn5Cre%sohiiR%tRccmsesaaCaeC(allssniIpsdeIhdss1sana1iPiia.atatys9.Ciyi9ggies8vts8Rnnnii(tvt(oFttooyaFosisistnig1idiy1gsosd7u7i)u)f.aor2.pwr2wegft9em9hne)t2e).2hero)P.re).TeesTC.SeTh1ThS1aRd4he4aeht).Dea.t2ses2D(sp9sNpT9s%eenNae%eAncbecnAi(siglfis(9feita9iitc5ettcit53iie%vsiitv%)tvstyi.yitteCtibwiCebwbeIysyaIsya6so6sp.opo3f.1fm1m3u8k00k8r0PDtP0Dor%tC%CNooN2RuRfA62fAo,to,.6i2rnracc.a22SaaneSn)2alldacdc)dattuuDniaDSaSlldnaNaagaNtdttnt7DAeeDA.od7d1NNsa.4a1inbAb%snA4daad%wsts(tekee9kees(Ddd5srD9tet%sN5soNoc%bnAbnCoAyynIcCtcitfla1ihliaiIhirn.rPr9n1gPmigC8i.ceC9ceeatRta8tRsdolsl analyzed by both polymerase chain reaction (PCR) tests (Table 3)

  • The overall sensitivities obtained in this study from both PCR assays are very low, suggesting that this approach of analysis cannot be considered a confirmatory test for clinic patients or blood donors, and the serology is still the first choice

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Summary

Introduction

Chagas disease had been described for the first time in 1909 by the Brazilian medical doctor Carlos Chagas. It is a neglected tropical disease (NTD) caused by the protozoa, single-celled eukaryotes, haemoflagellate Trypanosoma cruzi. It is transmitted mainly by a bed-bug insect called “kissing bugs” or “blood suckers” in endemic areas, a triatominae vector Triatoma spp., a subfamily of the Reduviidae [1]. The asymptomatic chronic stage is called “chronic indeterminate Chagas disease,” and it is characterized by very low parasitemia, frequently undetectable and interpreted as a fluctuant parasitic load [2,3]. The disability-adjusted life years (DALYs) caused by Chagas disease, globally is up to 10.8 [5,6]

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