Evaluation of Bacteria and Fungi DNA Abundance in Human Tissues.

Whereas targeted and shotgun sequencing approaches are both powerful in allowing the study of tissue-associated microbiota, the human: microorganism abundance ratios in tissues of interest will ultimately determine the most suitable sequencing approach. In addition, it is possible that the knowledge of the relative abundance of bacteria and fungi during a treatment course or in pathological conditions can be relevant in many medical conditions. Here, we present a qPCR-targeted approach to determine the absolute and relative amounts of bacteria and fungi and demonstrate their relative DNA abundance in nine different human tissue types for a total of 87 samples. In these tissues, fungi genomes are more abundant in stool and skin samples but have much lower levels in other tissues. Bacteria genomes prevail in stool, skin, oral swabs, saliva, and gastric fluids. These findings were confirmed by shotgun sequencing for stool and gastric fluids. This approach may contribute to a more comprehensive view of the human microbiota in targeted studies for assessing the abundance levels of microorganisms during disease treatment/progression and to indicate the most informative methods for studying microbial composition (shotgun versus targeted sequencing) for various samples types.

Rethinking Cervical Cancer Screening in Brazil Post COVID-19: A Global Opportunity to Adopt Higher Impact Strategies.

The World Health Organization global call to eliminate cervical cancer encourages countries to consider introducing or improving cervical cancer screening programs. Brazil's Unified Health System (SUS) is among the world's largest public health systems offering free cytology testing, follow-up colposcopy, and treatment. Yet, health care networks across the country have unequal infrastructure, human resources, equipment, and supplies resulting in uneven program performance and large disparities in cervical cancer incidence and mortality. An effective screening program needs multiple strategies feasible for each community's reality, facilitating coverage and follow-up adherence. Prioritizing those at highest risk with tests that better stratify risk will limit inefficiencies, improving program impact across different resource settings. Highly sensitive human papillomavirus (HPV)-DNA testing performs better than cytology and, with self-collection closer to homes and workplaces, improves access, even in remote regions. Molecular triage strategies like HPV genotyping can identify from the same self-collected sample, those at highest risk requiring follow-up. If proven acceptable, affordable, cost-effective, and efficient in the Brazilian context, these strategies would increase coverage while removing the need for speculum exams for routine screening and reducing follow-up visits. SUS could implement a nationwide organized program that accommodates heterogenous settings across Brazil, informing a variety of screening programs worldwide.

T. vaginalis in riverside women in Amazonia, Brazil: an experience using the EVALYN® BRUSH vaginal self-collection device.

INTRODUCTION: The challenges related to the diagnosis of sexually transmitted infections present more complex factors in remote and hard-to-reach areas. The use of self-collection devices that facilitate the obtaining of a biological sample with high quality for sensitive molecular tests have been examined. This study aimed to evaluate the performance and acceptance of the Evalyn® Brush (Rovers® Medical Devices) for detection of T. vaginalis among women living in the riverside communities of Amazonas, Brazil. METHODOLOGY: The study included 300 riverside women. They received instructions for self-collection, carried out the task, and then answered a questionnaire on the use of the device. T. vaginalis was detected by Polymerase Chain Reaction, using primers TVK3/TVK7. RESULTS: The mean age of the women was 35.8 years, and most of them presented low schooling, low income, agricultural activity and lived in a marital union. All samples were positive for human genomic DNA (100%) and the prevalence of T. vaginalis infection was 5.6% (n = 17). Of the 300 women, 293 (97.7%) indicated that they liked the use of the device, 287 (95.7%) reported having had no difficulty in handling it, 265 (88.3%) did not feel any type of discomfort and 228 (76%) said they preferred the self-collection to the collection made by the professional, mainly due to privacy and comfort. CONCLUSIONS: The Evalyn® Brush proved reliable as a device for the collection of biological samples for molecular analysis and was well-accepted by women. Its use can be indicated in remote and hard to reach places.

Real-time PCR versus conventional PCR for malaria parasite detection in low-grade parasitemia.

We have optimized a faster and cheaper real-time PCR and developed a conventional genus specific PCR based on 18S rRNA gene to detect malaria parasites in low-grade parasitemias. Additionally, we compared these PCRs to the OptiMAL-IT test. Since there is no consensus on choice of standard quantitative curve in real-time assays, we decided to investigate the performance of parasite DNA from three different sources: "genome", amplicon and plasmid. The amplicon curve showed the best efficiency in quantifying parasites. Both PCR assays detected 100% of the clinical samples tested; the sensitivity threshold was 0.5 parasite/mul and no PCR positive reaction occurred when malaria parasites were not present. Conversely, if OptiMAL-IT were employed for malaria diagnosis, 30% of false-negative results could be expected. We conclude that PCR assays have potential for detecting malaria parasites in asymptomatic infections, in evaluation of malaria vaccine molecule candidates, for screening blood donors, especially in endemic areas, or even in monitoring malaria therapy.

Métodos de triagem para malária, em doadores de sangue de área endêmica / Screening methods for malaria, in blood donors in endemic area

Relizou-se um estudo controlado para comparar a presença parasitemiaa e anticorpos antiplasmódios em doadores expostos ao risco de infecçäo malárica, segundo os critérios fixados nas Normas Técnicas em Hemoterapia, do Miníterio da Saúde. Eles estabelecem a rejeiçäo dos candidatos à doaçäo que apresentaram quadro febril há 30 dias e malária há 12 meses, ou que frequentaram área de malária há 6 meses. Foram estudados 395 candidatos incluídos nos critérios de rejeiçäo (expostos) e 383 candidatos aptos (controles), selecionados na triagem de doadores do Hemocentro de Manaus (AM). Em ambos os grupos, realizou-se a gota espessa e o teste da laranja acridina (QBC) para o diagnóstico parasitológico e a imunofluorescência indireta para a pesquisa de anticorpos IgM e IgG ao Plamodium vivax e falciparum. Observou-se diferença significante entre expostos e controles em relaçäo à presença de parasitemia, porém mäo para a presença de anticorpos antiplasmódios (P<0.05). O QBC foi mais sensível do que a gota espessa para detectar parasitemia e a concordância entre ambos foi de 0.66 ao Indice Kappa