T. cruzi OligoC-TesT: A Simplified and Standardized Polymerase Chain Reaction Format for Diagnosis of Chagas Disease

Chagas disease (American trypanosomiasis) is caused by the protozoan parasite Trypanosoma cruzi and represents a major public health problem in Latin America. Furthermore, growing human population movements extend the disease distribution to regions outside the South American continent. Accurate diagnosis is crucial in patient care and in preventing transmission through blood transfusion, organ transplantation, or vertical transmission from mother to child. Routine diagnosis of Trypanosoma cruzi infection generally is based on detection of the host's antibodies against the parasite. However, antibody detection tests are liable to specificity problems and are of limited use in assessing treatment outcome and congenital infections. The introduction of the polymerase chain reaction (PCR) to amplify specific DNA sequences opened promising diagnostic perspectives. Despite its reported high sensitivity and specificity, broad use of the PCR technique in diagnosis of Chagas disease is hampered by its complexity and the lack of any standardization. We here present the development and evaluation of the T. cruzi OligoC-TesT, a simple and standardized dipstick format for detection of PCR amplified T. cruzi DNA. The new tool is an important step towards simplified and standardized molecular diagnosis of Chagas disease

Sensitivities and specificities of the <i>T. cruzi</i> satDNA and kDNA OligoC-TesTs on 187 Chagas patients and 114 endemic and non-endemic controls from Chile, Argentina, Spain, Sudan and D.R. Congo.

<p>Notes: 95% CI = 95% confidence interval; Non-end. = Non-endemic; I = first repetition; II = second repetition; N.D. = not done due to limited number of test kits available;</p><p>^a All Chagas patients recruited at the Hospital Ramón y Cajal in Madrid were from Bolivian origin.</p

Analytical sensitivity and analytical specificity of the <i>T. cruzi</i> satDNA OligoC-TesT and kDNA OligoC-TesT on DNA from 6 <i>T. cruzi</i> and 3 <i>T. rangeli</i> reference strains.

<p>DTU = discrete typing unit, NA = not applicable.</p

Demographic characteristics of the participants recruited in the study and specifications of blood collections and reference tests.

<p>Notes: ND: not done; a: ELISA Chagas III, GrupoBios S.A., Chile; b: In house protocol with <i>T. cruzi</i> Tulahuen strain; c: Chagatest ELISA recombinant v. 4.0, Wiener Lab, Argentina; d: Chagatest HAI, Wiener Lab, Argentina; e: In house protocol with <i>T. cruzi</i> Dm28, MC & T strain; f: In house protocol with <i>T. cruzi</i> Dm28,MC & T strain.</p