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Simultaneous Detection of Mycobacterium tuberculosis and Mycobacterium avium Complex by a Multiplex PCR
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Souza, Mariana Quaresma de

; Busatto, Caroline

; Reis, Ana Julia

; Groll, Andrea von

; Silva, Pedro Eduardo Almeida da

; Ramis, Ivy Bastos

Brazilian Archives of Biology and Technology

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2023, Volume 66 elocation e23220262

Abstract: En | Text: En | PDF: En

Abstract Infection disease caused by nontuberculous mycobacteria (NTM) have been increasingly reported and often manifest with the same symptoms of tuberculosis (TB). The identification of the causative agent is fundamental for the determination of an appropriate therapy, since each species of mycobacteria requires a specific treatment. Thus, rapid and accurate tests to identify mycobacteria of medical interest, such as Mycobacterium tuberculosis complex (MTBC) and Mycobacterium avium complex (MAC), are necessary in the clinical routine. The present study evaluated an in-house multiplex PCR to detect, in reference and clinical strains, the genus Mycobacterium and also M. tuberculosis and MAC. To identify the Mycobacterium genus, M. tuberculosis and MAC, it was amplified a fragment of hsp65 gene, esat-6 gene, and the internal transcribed spacer between the 16S and 23S rRNA genes, respectively. In total, 87 mycobacteria strains were used, being 10 reference and 77 clinical strains, previously identified as MTBC (n = 66), M. avium (n = 8) or other NTM specie (n = 13). The hsp65 gene fragment was amplified for all mycobacteria strains evaluated (87/87). This multiplex PCR presented sensitivity of 100% and specificity of 95.2% for M. tuberculosis detection, and sensitivity of 100% and specificity of 100% for MAC detection. The multiplex PCR evaluated is an important tool for the differentiation between M. tuberculosis and NTM, as well as for the identification of MAC, a complex composed by species with high prevalence in the world.

https://doi.org/10.1590/1678-4324-2023220262

Thesis and dissertations examining tuberculosis in Brazil between 2013 and 2019: an overview
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Reis, Ana Júlia

; Pizzol, Juliana Lemos Dal

; Gattelli, Rúbia

; Groll, Andrea von

; Ramos, Daniela Fernandes

; Ramis, Ivy Bastos

; Kritski, Afrânio

; Silva, José Roberto Lapa e

; Silva, Pedro Eduardo Almeida da

Revista da Sociedade Brasileira de Medicina Tropical

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2022, Volume 55 elocation e0198-2022

Abstract: En | Text: En | PDF: En

ABSTRACT Background: Tuberculosis (TB) remains a serious public health problem, with approximately 10 million new cases reported annually. Knowledge about the quantitative evolution of theses and dissertations (T&Ds) examining human TB in Brazil can contribute to generating strategic planning for training professionals in this field and disease control. Therefore, this study highlights the role of T&Ds on TB in national scientific disclosures. Methods: An integrative review related to TB was performed, including T&Ds produced in Brazil and completed between 2013 and 2019. Results: A total of 559,457 T&Ds were produced, of which 1,342 were associated with TB, accounting for 0.24% of the total number of T&Ds in Brazil. This was evidenced by a predominance of themes such as attention/health care, epidemiology, and TB treatment, and 80.2% of the T&Ds on TB were related to the large areas of health and biological sciences. Only 19.7% of T&Ds were associated with groups of patients considered at risk for TB, and 50.9% were produced in southeastern Brazil. The 1,342 T&Ds on TB were developed in 416 postgraduate programs linked to 121 higher education institutions (HEIs). We highlight that 72.7% of T&Ds on TB were produced in federal HEIs, 27.4% in state HEIs, and 8.5% in private HEIs. Conclusions: Strategic themes, such as TB control, require public policies that aim to increase the number of doctors and masters with expertise in TB, with geographic uniformity, and in line with the priorities for disease control.

https://doi.org/10.1590/0037-8682-0198-2022

Protective effect of the probiotic Lactobacillus acidophilus ATCC 4356 in BALB/c mice infected with Toxocara canis
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Cadore, Priscila Silva

; Walcher, Débora Liliane

; Sousa, Nicholas Frota Gonçalves Correia de

; Martins, Lourdes Helena Rodrigues

; Hora, Vanusa Pousada da

; Groll, Andrea Von

; Moura, Micaele Quintana de

; Berne, Maria Elisabeth Aires

; Avila, Luciana Farias da Costa de

; Scaini, Carlos James

Revista do Instituto de Medicina Tropical de São Paulo

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2021, Volume 63 elocation e9

Abstract: En | Text: En | PDF: En

ABSTRACT Human toxocariasis consists of chronic tissue parasitosis that is difficult to treat and control. This study aimed to evaluate the action of the probiotic Lactobacillus acidophilus ATCC 4356 on larvae of Toxocara canis and the effect of IFN-γ cytokine on parasite-host in vivo (1.109 CFU) and in vitro (1.106, 1.107, 1.108, 1.109 CFU) interactions. Four groups of six BALB/c mice were formed: G1 - L. acidophilus supplementation and T. canis infection; G2 - T. canis infection; G3 - L. acidophilus supplementation; and G4 - PBS administration. Mice were intragastrically suplemented with probiotics for 15 days before inoculation and 48 h after inoculation with 100 T. canis eggs. The inoculation of T. canis was also perfomed intragastrically. The recovery of larvae took place through digestion of liver and lung tissues; the evaluation of IFN-γ gene transcription in leukocytes was performed by qPCR. The in vitro test consisted of incubating the probiotic with T. canis larvae. The supplementation of probiotics produced a reduction of 57.7% (p = 0.025) in the intensity of infection of T. canis larvae in mice, whereas in the in vitro test, there was no larvicidal effect. In addition, a decrease in the IFN-γ gene transcription was observed in both, T. canis-infected and uninfected mice, regardless of whether or not they received supplementation. The probiotic L. acidophilus ATCC 4356 reduced T. canis infection intensity in mice, however, the probiotic did not have a direct effect on larvae, demonstrating the need of interaction with the host for the beneficial effect of the probiotic to occur. Yet, the proinflammatory cytokine IFN-γ did not apparently contributed to the observed beneficial effect of probiotics.

https://doi.org/10.1590/s1678-9946202163009 656 downloads

Molecular detection of Paracoccidioides in soil from an urban area of southern Brazil
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Mendes, Josiara Furtado

; Poester, Vanice Rodrigues

; Groll, Andrea Von

; Meireles, Mário Carlos Araújo

; Xavier, Melissa Orzechowski

Revista da Sociedade Brasileira de Medicina Tropical

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2020, Volume 53 elocation e20190172

Abstract: En | Text: En | PDF: En

Abstract INTRODUCTION: Previous studies that detected Paracoccidioides spp. DNA in soil taken from rural areas have shown this to be a valuable tool for ecological and epidemiological studies. This study reports the detection of Paracoccidioides spp. DNA in soil samples from an urban area of southern Brazil. METHODS: Sixteen soil samples were submitted to nested-PCR and the amplicons of a representative number of positive samples were sequenced. RESULTS: Paracoccidioides spp. DNA was found in 44% of samples. Four DNA amplicons were sequenced, showing 100% homology with P. brasiliensis. CONCLUSIONS: The southern Brazilian urban population is commonly exposed to the Paracoccidioides fungus.

https://doi.org/10.1590/0037-8682-0172-2019 382 downloads

Laboratory diagnosis for Covid-19: A mini-review
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Pizzol, Juliana Lemos Dal

; Hora, Vanusa Pousada da

; Reis, Ana Júlia

; Vianna, Júlia

; Ramis, Ivy

; Groll, Andrea von

; Silva, Pedro Almeida da

Revista da Sociedade Brasileira de Medicina Tropical

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2020, Volume 53 elocation e20200451

Abstract: En | Text: En | PDF: En

Abstract Coronavirus disease (COVID-19) is a pandemic caused by a new coronavirus, called SARS-CoV-2. This disease was first identified in December 2019 and rapidly developed into a challenge to the public health systems around the world. In the absence of a vaccine and specific therapies, disease control and promotion of patient health are strongly dependent on a rapid and accurate diagnosis. This review describes the main laboratory approaches to making a diagnosis of COVID-19 and identifying those previously infected with SARS-CoV-2.

https://doi.org/10.1590/0037-8682-0451-2020 4856 downloads

Detection of Drug Resistant Mycobacterium Tuberculosis Strains Using Kit SIRE Nitratase®: a Multicenter Study
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Miranda, Silvana Spindola de

; Almeida, Isabela Neves de

; Mansur, Maria de Fátima Filardi Oliveira

; Figueredo, Lida Jouca de Assis

; Carvalho, Wânia da Silva

; Hadaad, João Paulo Amaral

; Diniz, Jaciara de Lourdes do Carmo Guimarães

; Groll, Andrea von

; Silva, Pedro Almeida da

; Lopes, Maria Luiza

; Santos, Marcelo Cordeiro dos

; Brito, Alexandra

; Mello, Fernanda Carvalho de Queiroz

; Malaquias, Thiago da Silva Santos

; Croda, Julio

; Pinhata, Juliana Maira Watanabe

; Oliveira, Rosângela Siqueira de

; Chimara, Erica

; Rossetti, Maria Lúcia

; Halon, Maria Laura

; Lourenço, Maria Cristina

; Medeiros, Reginalda Ferreira de Melo

; Montes, Fátima Cristina Onofre Fandinho

; Machado, Diana

; Viveiros, Miguel

; Kritski, Afrânio Lineu

Brazilian Archives of Biology and Technology

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2020, Volume 63 elocation e20190179

Abstract: En | Text: En | PDF: En

Abstract (1) Background: The Commercial Kit SIRE Nitratase® PlastLabor, is a drug susceptibility test kit used to detect Mycobacterium tuberculosis resistance to first-line TB treatment drugs. The present study aimed at evaluating its performance in a multicenter study. (2) Methods: To determine its accuracy, the proportion methods in Lowenstein Jensen medium or the BACTECTMMGITTM960 system was used as a gold standard. (3) Results: The study revealed that the respective accuracies of the kit with 190 M. tuberculosis clinical isolates, using the proportion methods in Lowenstein Jensen medium or BACTECTMMGITTM960 system as a gold standard, were 93.9% and 94.6%, 96.9% and 94.6%, 98.0% and 97.8%, and 98.0% and 98.9%, for streptomycin, isoniazid, rifampicin, and ethambutol, respectively. (4) Conclusion: Thus, the kit can rapidly screen resistance to streptomycin, isoniazid, rifampicin, and ethambutol. Additionally, it does not require sophisticated equipment; hence, it can be easily used in the laboratories of low and middle income countries.

https://doi.org/10.1590/1678-4324-2020190179 905 downloads

Nontuberculous mycobacteria in patients with suspected tuberculosis and the genetic diversity of Mycobacterium avium in the extreme south of Brazil
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Busatto, Caroline

; Vianna, Júlia Silveira

; Silva, Ana Barbara Scholante

; Basso, Rossana

; Silveira, Jussara

; Groll, Andrea Von

; Ramis, Ivy Bastos

; Silva, Pedro Eduardo Almeida da

Jornal Brasileiro de Pneumologia

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2020, Volume 46 N. 2 elocation e20190184

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO Objetivo: As micobactérias não tuberculosas (MNT) são um grupo heterogêneo de bactérias amplamente distribuídas na natureza e relacionadas com infecções oportunistas em seres humanos. Os objetivos deste estudo foram identificar MNT em pacientes com suspeita de tuberculose e culturas positivas e avaliar a diversidade genética de cepas identificadas como Mycobacterium avium. Métodos: Foram estudadas amostras pulmonares e extrapulmonares provenientes de 1.248 pacientes. As amostras que apresentaram resultado positivo em cultura e negativo para o complexo M. tuberculosis na identificação molecular foram avaliadas por meio da detecção dos genes hsp65 e rpoB e de sequenciamento de fragmentos conservados desses genes. Todas as cepas identificadas como M. avium foram genotipadas pelo método mycobacterial interspersed repetitive unit-variable-number tandem-repeat com oito loci. Resultados: Das 332 micobactérias isoladas, 25 (7,5%) eram MNT. Dessas 25, 18 (72%) eram M. avium, 5 (20%) eram M. abscessus, 1 (4%) era M. gastri e 1 (4%) era M. kansasii. As 18 cepas de M. avium apresentaram alta diversidade, e apenas duas eram geneticamente relacionadas. Conclusões: Esses resultados mostram a necessidade de considerar a investigação de MNT em pacientes com suspeita de tuberculose ativa e culturas positivas e de avaliar a diversidade genética de cepas de M. avium.

ABSTRACT Objective: Nontuberculous mycobacteria (NTM) are a heterogeneous group of bacteria that are widely distributed in nature and associated with opportunistic infections in humans. The aims of this study were to identify NTM in patients with suspected tuberculosis who presented positive cultures and to evaluate the genetic diversity of strains identified as Mycobacterium avium. Methods: We studied pulmonary and extrapulmonary samples obtained from 1,248 patients. The samples that tested positive on culture and negative for the M. tuberculosis complex by molecular identification techniques were evaluated by detection of the hsp65 and rpoB genes and sequencing of conserved fragments of these genes. All strains identified as M. avium were genotyped using the eight-locus mycobacterial interspersed repetitive unit-variable-number tandem-repeat method. Results: We found that NTM accounted for 25 (7.5%) of the 332 mycobacteria isolated. Of those 25, 18 (72%) were M. avium, 5 (20%) were M. abscessus, 1 (4%) was M. gastri, and 1 (4%) was M. kansasii. The 18 M. avium strains showed high diversity, only two strains being genetically related. Conclusions: These results highlight the need to consider the investigation of NTM in patients with suspected active tuberculosis who present with positive cultures, as well as to evaluate the genetic diversity of M. avium strains.

https://doi.org/10.36416/1806-3756/e20190184 734 downloads

Sporothrix spp. EVALUATION IN SOIL OF A HYPERENDENIC AREA FOR SPOROTRICHOSIS IN SOUTHERN BRAZIL
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Poester, Vanice Rodrigues

; Mendes, Josiara Furtado

; Groll, Andrea Von

; Klafke, Gabriel Baracy

; Brandolt, Tchana Martinez

; Xavier, Melissa Orzechowski

Ciência Animal Brasileira

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2018, Volume 19 elocation e-52571

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo O aumento de casos de esporotricose zoonótica tornou-se um problema de saúde pública em áreas hiperendêmicas para a doença em felinos. O envolvimento dos gatos domésticos nesta transmissão e o contato contínuo destes animais com o solo sugerem que o ambiente possa ser uma importante fonte e/ou reservatório de fungos Sporothrix spp. Assim, este estudo objetivou avaliar a presença de fungos do complexo Sporothrix em amostras de solo provenientes de locais de acesso de felinos infectados no sul do Brasil. Foram coletadas amostras de solo de residências com felinos com esporotricose, de locais próximos a estas residências e também de locais públicos, totalizando 101 amostras. Estas foram processadas pela técnica de plaqueamento direto e incubadas a 25°C por até 15 dias. Dezessete amostras foram paralelamente submetidas a extração de DNA (kitNORGEN BIOTEK CORP®) e técnica de PCR espécie-específico e Nested-PCR. Embora em nenhuma amostra avaliada tenha sido detectada a presença de Sporothrix spp., seja por cultivo ou por biologia molecular, este estudo não descarta o papel do ambiente na tríade do processo infeccioso (solo - animal - humano), sendo necessários outros estudos ampliando área de abrangência, volume, tipo e método de coleta das amostras, bem como técnicas de detecção.

Abstract The increase in zoonotic sporotrichosis cases has become a public health problem in hyperendemic areas for the feline disease. Domestic cats involvement in this transmission and the continuous contact of these animals with soil suggest that the environment may be an essential source and/or reservoir of Sporothrix spp. Thus, the current study aimed to evaluate the presence of Sporothrix spp. in the soil of places where infected cats in Southern Brazil is present. In total, 101 soil samples were collected from the residence of cats with sporotrichosis, from regions around to these residences, and also from public places. These samples were processed using the direct plating technique being incubated at 25 °C for up to 15 days. Additionally, seventeen samples were also submitted to DNA extraction (NORGEN BIOTEK CORP® kit), PCR-specific and Nested-PCR technique. Although the presence of Sporothrix in soil has been undetected, by culture or molecular biology, this study does not reject the role of the environment in the triad of the infectious process (soil-animal-human). Thus, further studies are necessary for expanding the area of coverage, volume, type and method of sampling, and/or detection techniques.

https://doi.org/10.1590/1809-6891v19e-52571 2157 downloads

Authors reply: cagE as a biomarker of the pathogenicity of Helicobacter pylori
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Ramis, Ivy Bastos

; Vianna, Júlia Silveira

; Silva Junior, Lande Vieira da

; Groll, Andrea Von

; Silva, Pedro Eduardo Almeida da

Revista da Sociedade Brasileira de Medicina Tropical

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jun 2014, Volume 47 N. 3 Pages 404 - 404

Abstract: En | Text: En | PDF: En

https://doi.org/10.1590/0037-8682-0261-2013 1015 downloads

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Nitrate reductase assay using sodium nitrate for rapid detection of multidrug resistant tuberculosis
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Macedo, Maíra Bidart

; Groll, Andrea Von

; Fissette, Krista

; Palomino, Juan Carlos

; Silva, Pedro Eduardo Almeida da

; Martin, Anandi

Brazilian Journal of Microbiology

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sep 2012, Volume 43 N. 3 Pages 981 - 983

Abstract: En | Text: En | PDF: En

We validated the nitrate reductase assay (NRA) for the detection of multidrug-resistant Mycobacterium tuberculosis (MDR-TB) using sodium nitrate (NaNO3) in replacement of potassium nitrate (KNO3) as nitrate source. NaNO3 is cheaper than KNO3 and has no restriction on use which facilitates the implementation of NRA to detect MDR-TB.

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