DOI: 10.5061/dryad.05qfttf8f
title: Data from: Quantitative PCR as a marker for preemptive therapy and its role in therapeutic control in Trypanosoma cruzi/HIV coinfection

description: Background: Trypanosoma cruzi and HIV coinfection can evolve with
 depression of cellular immunity and increased parasitemia. We applied
 quantitative PCR (qPCR) as a marker for preemptive antiparasitic treatment
 to avoid fatal Chagas disease reactivation and analyzed the outcome of
 treated cases. Methodology: This mixed cross-sectional and longitudinal
 study included 171 Chagas disease patients, 60 coinfected with HIV. Of
 these 60 patients, ten showed Chagas disease reactivation, confirmed by
 parasites identified in the blood, cerebrospinal fluid, or tissues, 12
 exhibited high parasitemia but no reactivation, and 38 had low parasitemia
 and no reactivation. Results:  We showed, for the first time, the
 success of the timely introduction of benznidazole in the non-reactivated
 group with high levels of parasitemia detected by qPCR and the absence of
 parasites in reactivated cases with at least 58 days of benznidazole. HIV+
 and HIV+ without reactivation had a 4.0 – 5.1 higher chance of having
 parasitemia than HIV seronegative cases. A positive correlation was found
 between parasite and viral loads. Remarkably, treated T. 
 cruzi/HIV-coinfected patients had 77.3% conversion from positive to
 negative parasitemia compared to 19.1% of untreated patients.
 Additionally, untreated patients showed ~13.6 times higher odds of having
 positive parasitemia in the follow-up period compared with treated
 patients. Treated and untreated patients showed no differences regarding
 the evolution of Chagas disease. The main factors associated with
 all-cause mortality were higher parasitemia, lower CD4 counts/µL, higher
 viral load, and absence of antiretroviral therapy. Conclusion: We
 recommend qPCR prospective monitoring of T. cruzi parasitemia in HIV+
 patients and point out the value of pre-emptive therapy for patients with
 temporary high parasitemia. In parallel, an early antiretroviral therapy
 introduction is advisable, aiming at viral load control, immune response
 restoration, and major survival. We also suggest an earlier
 antiparasitic treatment for all coinfected patients, followed by
 effectiveness analysis alongside antiretroviral therapy.
description: The mixed cross-sectional and longitudinal study included 171
 patients with <em>T. cruzi </em>infection: 60 HIV-coinfected
 (HIV+) and 111 HIV seronegative, as comparator groups. Most of them
 recruited from the Infectious and Parasitic Diseases Division of the
 Hospital das Clínicas, a tertiary hospital adjacent to the School of
 Medicine of the University of São Paulo, Brazil. All sociodemographic,
 clinical, and laboratory data, date of treatment and sample collections
 (before treatment and during follow-up), results of parasitemia
 (parasitological and molecular methods), CD4 counts, HIV viral loads,
 antiretroviral therapy and schemes, the evolution of Chagas disease,
 all-cause mortality and deaths due to Chagas disease, were collected by
 accessing the Medical records.
description: # Data from: Quantitative PCR as a marker for preemptive therapy and its
 role in therapeutic control in Trypanosoma cruzi//HIV coinfection.
 [https://doi.org/10.5061/dryad.05qfttf8f](https://doi.org/10.5061/dryad.05qfttf8f) A positive correlation was found between parasites and viral loads. Remarkably, treated *T.  cruzi/*HIV-coinfected patients had 77.3% conversion from positive to negative parasitemia compared to 19.1% of untreated patients. In addition, untreated patients showed ~13.6 times higher odds of having positive parasitemia in the follow-up period compared with treated patients. There were no statistically significant differences between treated and untreated patients regarding the evolution of Chagas disease. The main factors associated with all-cause deaths were higher parasitemia, lower CD4 counts/µL, higher viral load, and absence of antiretroviral therapy. ## Description of the data and file structure The study included 171 patients with *T. cruzi *infection: 60 HIV-coinfected (HIV+) and 111 HIV seronegative, as comparator groups. All sociodemographic, clinical, and laboratory data, date of therapy and, sample collections (before treatment and during follow-up), results of parasitemia (parasitological and molecular methods), CD4 counts, HIV viral loads, antiretroviral therapy and schemes, the evolution of Chagas disease, all-cause mortality and deaths due to Chagas disease, were collected by accessing the Medical records, and organized in an excel spreadsheet. The meaning of the variables is described in the following table: | Row | Definition | | :-- | :------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | | A | Identification | | B | Chagas Disease patients with (Yes) or witlhout  co-infection HIV (No) | | C | Native to which region of Brazil | | D | Sex; Male=0, Female=1 | | E | Missing - Information not available | | F | 2 - Information not available | | G | Cardiac Form of CD: Cardiac involvement and Cardiac + Digestive involvement; 2 - Information not available | | H | Digestive Form of CD: Digestive involvement and Digestive + Cardiac involvement; 2 - Information not available | | I | Indeterminated Form of CD: Patients without signals or symptoms and without Electrocardiogram changes, normal cardiac area and no digestive involvement; 2 - Information not available | | J | Date in quarter/year of the 1st sample collected intended for testing to detect T. cruzi using direct parasitological (QBC; microhematocrit, histopathology), indirect parasitological (blood culture and xenodiagnosis), and molecular tests (conventional and quantitative PCR) and/or 1st clinical evaluation of the patient. | | K | Reactivation of chronic Chagas disease is a rare condition and occurs only in immunosuppressed patients.  This condition is diagnosed by the detection of T. cruzi by direct parasitological tests: QBC, Microhematocrit, Strout, histopathological. | | L | Site of manifestation of Chagas disease reactivation. | | M | cPCR T0: Conventional PCR referring to the 1st sample collected. Results presented as Positive or Negative. | | N | qPCR, Par Eq/ml of blood, T0, (Log10): Quantitative PCR, results presented as log10 (equivalent value of T. cruzi/mL of blood collected +1), referring to the 1st sample collected. | | O | Indirect Parasitilogical Test: blood culture and xenodiagnosis results of 1st sample collected. Results presented as Positive or Negative | | P | Parasitemia: parasitological indirect (blood culture and xenodiagnosis) and/or molecular tests (conventional and quantitative PCR, referring to the 1st sample collected. Results presented as Positive or Negative | | Q | Patients with reactivation of Chagas disease or those not reactivated who presented high levels of parasitemia, assessed by quantitative xenodiagnosis, associated with clinical worsening of Chagas disease were treated with Benzonidazole. | | R | CD4\* count cells/µL T0 = T CD4 cell count in HIV+ patients. Results presented as number of cells/ul | | S | Viral Load count RNA viral copies\*/µL T0 = RNA viral copies in HIV+ patients. Results presented as number of RNA viral copies\*/µL. | | T | ART- T0: Use of antiretroviral therapy before and during the collection of the first sample | | U | ART class: Class of antiretroviral therapy used | | V | Date in quarter/year of the 2nd sample collected intended for testing to detect T. cruzi using direct parasitological tests (QBC; microhematocrit, histopathology), indirect (blood culture and xenodiagnosis), and molecular tests (conventional and quantitative PCR) | | W | cPCR T1:Conventional  PCR referring to the 2nd sample collected. Results presented as Positive or Negative | | X | qPCR, Par Eq/ml of blood, T1, (Log10): Quantitative PCR, results presented as log10 (equivalent value of T cruzi/mL of blood collected +1), referring to the 2nd sample collected | | Y | Indirect Parasitilogical Test: blood culture and xenodiagnosis results of 2nd sample collected. Results presented as Positive or Negative | | Z | Parasitemia: parasitological indirect (blood culture and xenodiagnosis) and/or molecular tests (conventional and quantitative PCR), referring to the 2nd sample collected. Results presented as Positive or Negative | | AA | CD4\* count cells/µL T0 = T CD4 cell count in HIV+ patients,  referring to the 2nd sample collected. Results presented as number of cells/ul | | AB | Viral Load count RNA viral copies\*/µL T0 = RNA viral copies in HIV+ patients,  referring to the 2nd sample collected. Results presented as number of RNA viral copies\*/µL. | | AC | CD Clinical Worsening:  Clinical worsening of Chagas disease during follow-up is characterized by evolution to more severe disease in the same clinical form or evolution to another clinical form or death.  | | AD | Non-immunosuppressed patients with chronic Chagas disease were referred for follow-up in basic health units, which leads to loss of information. | | AE | Date in quarter/year of the dead.  | ## Change log: October 1, 2025: Added Human Subjects Data section to README ## Human subjects data I confirm that I have received explicit consent from participants to publish the data. The consent form explicitly guarantees confidentiality. Below is the English version of the consent form that was used. ANEXO I HOSPITAL DAS CLÍNICAS DA FACULDADE DE MEDICINA DA UNIVERSIDADE DE SÃO PAULO-HCFMUSP FREE INFORMED CONSENT FORM _________________________________________________________________ IDENTIFICATION DATA OF THE SUBJECT OF THE RESEARCH OR LEGAL GUARDIAN 1. Name: .:............................................................................. ........................................................... IDENTIFICATION DOCUMENT Nº : ........................................ SEX : M □ F □ DATE OF BIRTH: ......../......../...... ADDRESS ............................................................................................................... Nº ................................... CITY/STATE ............................................................. ZIP CODE:......................................... FONE: DDD (............) .......................................................................... 2. LEGAL GUARDIAN.................................................................................................................................. TYPE (degree of kinship, guardian, curator, etc.) .................................................................................. IDENTIFICATION DOCUMENT Nº:....................................SEX: M □ F □ DATE OF BIRTH: ......../......../...... ADDRESS ............................................................................................................... Nº ................................... CITY/STATE ............................................................. ZIP CODE:......................................... FONE: DDD (............) .......................................................................... ________________________________________________________________________________________________ RESEARCH DATA TITLE: Monitoring of immunocompromised patients with Chagas disease: parasitemia levels and association with profile and cytokine polymorphism, survival, and characteristics of Trypanosoma cruzi isolates in organs transplants, use of immunosuppressive drugs, and co-infection with human immunodeficiency vírus PRINCIPAL INVESTIGATOR: Profa. Maria Aparecida Shikanai Yasuda POSITION/FUNCTION: Full Professor REGIONAL MEDICAL COUNCIL REGISTRATION. State of São Paulo. No.: 14.500 HCFMUSP UNIT: Department of Infectious and Parasitic Diseases 3. RESEARCH RISK ASSESSMENT: NO RISK □ MINIMAL RISK □x MEDIUM RISK □ LOW RISK □ HIGH RISK □ 4. RESEARCH DURATION: 24 months HOSPITAL DAS CLÍNICAS DA FACULDADE DE MEDICINA DA UNIVERSIDADE DE SÃO PAULO-HCFMUSP 1 – Study design and objective(s): In addition to verifying the differences between the number of microbes (parasites) that cause Chagas disease, this study aims to study the natural response of patients to infection. To this end, the DNA of the parasite and the patient will be analyzed in the laboratory. The levels of components of your immune system will also be studied in your serum. The results may help to clarify why some patients have heart, esophagus, and intestinal involvement and others do not. In the future, the results may be useful for the control and treatment of Chagas disease. Tests will be performed to assess the function of your heart and part of your digestive tract. A three-dimensional ultrasound-like exam may also be performed, which is minimally risky and provides more details about the heart. No problems have been reported from performing this exam, and there are no contraindications for performing it. An assessment of your quality of life will also be performed using a questionnaire. 2 – Description of the procedures that will be performed, their purposes, and identification of those that are experimental and not routine: You will be monitored for at least 1 year, with serological, parasitological, and molecular tests, and a small sample will be saved for further studies related to this research. To perform these tests, 10 mL of blood will be collected from a vein in your forearm. The frequency will depend on the type of disease and/or transplant you have. In patients with reactivation of Chagas disease or with a large number of parasites in the blood, specific treatment with benznidazole will be proposed. 3 – List of routine procedures and how they are performed: Blood collection through peripheral puncture of the forearm vein. 4 – Description of expected discomfort and risks in procedures 2 and 3: Blood collection may cause pain, bruising, and minor bleeding at the site. 5 – Benefits for the participant: Although there is no direct benefit for you from participating in the study, your contribution will be valuable in helping us better understand the best time to treat Chagas disease in patients undergoing transplantation and/or in patients using medications that weaken the immune system. In addition, through this study, there is a possibility of finding a marker for the most severe forms of the disease. HOSPITAL DAS CLÍNICAS DA FACULDADE DE MEDICINA DA UNIVERSIDADE DE SÃO PAULO-HCFMUSP 6 – List of alternative procedures that may be beneficial, which the patient may choose: None. 7 – Guaranteed access: At any stage of the study, you will have access to the professionals responsible for the research to clarify any questions you may have. The principal investigator is Dr. Maria Aparecida Shikanai Yasuda, who can be found at the following address: Av. Dr. Enéas de Carvalho Aguiar, 255, 4th floor - MI - room 4028, Division of Infectious and Parasitic Diseases, tel. 3069-6530 to answer questions. If you have any comments or questions about research ethics, please contact the Research Ethics Committee (CEP) – Rua Ovídio Pires de Campos, 225 – 5th floor – tel: 3069-6442 extensions 16, 17, 18 or 20, FAX: 3069-6442 extension 26 – E-mail: cappesq@hcnet.usp.br 8 – You are guaranteed the freedom to withdraw your consent at any time and to stop participating in the study, without any prejudice to the continuity of your treatment at the Institution; 9 – Right to confidentiality: The information obtained will be analyzed in conjunction with other patients, and the identification of any patient will not be disclosed; 10 – Right to be kept up to date on the partial results of the research, in open studies, or on results that are known to the researchers; 11 – Expenses and compensation: There are no personal expenses for the participant at any stage of the study, including tests and consultations. There is also no financial compensation related to their participation. If there are any additional expenses, they will be absorbed by the research budget. 12 – Commitment by the researcher to use the data and material collected only for this research. Explanation of compensation guarantee in the event of any damages resulting from the research. You will be guaranteed compensation for any damages resulting from blood collection during the research, in accordance with current legislation. HOSPITAL DAS CLÍNICAS DA FACULDADE DE MEDICINA DA UNIVERSIDADE DE SÃO PAULO-HCFMUSP I believe I have been sufficiently informed about the information I have read or that has been read to me, describing the study “Monitoring of immunocompromised patients with Chagas disease: parasitemia levels and association with profile, cytokine polymorphism, survival, and characteristics of Trypanosoma cruzi isolates in organ transplantation, use of immunosuppressive drugs, and co-infection with human immunodeficiency virus." I discussed my decision to participate in this study with Dr. Maria Aparecida Shikanai Yasuda. The purposes of the study, the procedures to be performed, associated discomforts and risks, the guarantees of confidentiality and ongoing clarification were made clear to me. It was also explained that my participation is free of charge and that I am guaranteed access to hospital treatment when necessary. I voluntarily agree to participate in this study and may withdraw my consent at any time, before or during the study, without penalty or prejudice or loss of any benefits I may have acquired, or in my care at this Service. ------------------------------------------------- Patient/legal representative signature Date / / ------------------------------------------------------------------------- Witness signature* Date / / (*For cases involving patients under the age of 18, patients who are illiterate or semi-literate, and patients with hearing or visual impairments). (For the project manager only) I declare that I have obtained the informed consent of this patient or legal representative for participation in this study in an appropriate and voluntary manner. ------------------------------------------------------------------------- Signature of the person responsible for the study Data / /