Pesquisa | SciELO

BACKGROUND Chikungunya is a mosquito-borne virus that has been causing large outbreaks in the Americas since 2014. In Brazil, Asian-Caribbean (AC) and East-Central-South-African (ECSA) genotypes have been detected and lead to large outbreaks in several Brazilian states. In Rio Grande do Sul (RS), the southernmost state of Brazil, the first cases were reported in 2016. OBJECTIVES AND METHODS We employed genome sequencing and epidemiological investigation to characterise the Chikungunya fever (CHIKF) burden in RS between 2017-2021. FINDINGS We detected an increasing CHIKF burden linked to travel associated introductions and communitary transmission of distinct lineages of the ECSA genotype during this period. MAIN CONCLUSIONS Until 2020, CHIKV introductions were most travel associated and transmission was limited. Then, in 2021, the largest outbreak occurred in the state associated with the introduction of a new ECSA lineage. CHIKV outbreaks are likely to occur in the near future due to abundant competent vectors and a susceptible population, exposing more than 11 million inhabitants to an increasing infection risk. mosquitoborne mosquito borne 2014 Brazil AsianCaribbean Asian Caribbean AC (AC EastCentralSouthAfrican East Central South African (ECSA states RS, , (RS) 2016 (CHIKF 20172021. 20172021 2017 2021. 2021 2017-2021 period 2020 limited Then lineage population 1 risk 201 (RS 2017202 202 2017-202 20 201720 2017-20 2 20172 2017-2 2017-

https://doi.org/10.1590/0074-02760220259

Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA
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Morais, Franciele Costa Leite

; Bello, Graziele Lima

; Costi, Cíntia

; Schmid, Karen Barros

; Soares, Tainá dos Santos

; Barcellos, Regina Bones

; Unis, Gisela

; Dias, Claudia Fontoura

; da Silva, Pedro Eduardo Almeida

; Rossetti, Maria Lucia

Memórias do Instituto Oswaldo Cruz

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2022, Volume 117 elocation e220031

Resumo: En | Texto: En | PDF: En

BACKGROUND Non-tuberculous mycobacteria (NTMs) cause diseases known as mycobacteriosis and are an important cause of morbidity and mortality. The diagnosis of pulmonary disease caused by NTM is hampered by its clinical similarity with tuberculosis (TB) and by the lack of an accurate and rapid laboratory diagnosis. OBJECTIVES Detect DNA from NTMs directly from lung samples using real-time polymerase chain reaction (qPCR) for amplification of 16S rRNA. Additionally, DNA sequencing (hsp65 and rpoB genes) was used to identify the species of MNTs. METHODS A total of 68 sputum samples (54 with suspected NTMs and 14 with TB) from patients treated at a referral hospital were used. FINDINGS Of these, 27/54 (50%) were qPCR positive for NTMs and 14/14 TB patients (controls) were qPCR negative with an almost perfect concordance (Kappa of 0.93) with the Mycobacterium spp. culture. Sequencing confirmed the presence of NTM in all positive samples. The most common species was Mycobacterium gordonae (33%), followed by Mycobacterium abscessus (26%), Mycobacterium fortuitum (22%), Mycobacterium avium (15%) and Mycobacterium peregrinum (4%). MAIN CONCLUSIONS The qPCR technique for detecting NTMs targeting 16S rRNA has the potential to detect NTMs and rapidly differentiate from Mycobacterium tuberculosis. However, it is necessary to identify the species to help in the differential diagnosis between disease and contamination, and to guide the choice of the therapeutic scheme.

https://doi.org/10.1590/0074-02760220031

Ética Algorítmica: questões e desafios éticos do avanço tecnológico da sociedade da informação
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Rossetti, Regina

; Angeluci, Alan

Galáxia (São Paulo)

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2021, Nº 46 elocation e50301

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

Resumo A Quarta Revolução Industrial inseriu na vida cotidiana a Inteligência Artificial (IA) e seus algoritmos. Essa nova realidade tecnológica requer uma profunda reflexão ética e enseja a necessidade de pensar sobre uma Ética Algorítmica. Partindo de uma pesquisa bibliográfica, este artigo propõe uma definição do termo e desenvolve uma reflexão sobre as principais questões suscitadas pela presença dos algoritmos nas mediações da vida humana contemporânea, referentes à falibilidade, opacidade, viés, discriminação, autonomia, privacidade e responsabilidade. Situando-a no domínio de uma ética aplicada, pondera-se sobre a conjunção das camadas humanas e computacionais na tomada de consciência diante dos dilemas.

Abstract The Fourth Industrial Revolution has put Artificial Intelligence (AI) and its algorithms into everyday life. This new technological reality requires a deep ethical reflection and gives rise to the need of thinking about an Algorithmic Ethic. From a bibliographic research, this article proposes a definition of the ethics of the algorithms, presenting a reflection on the main ethical issues raised by the presence of the algorithms in the mediations of contemporary human life regarding fallibility, opacity, bias, discrimination, autonomy, privacy and responsibility. Situating it in the domain of an applied ethic, one ponders about the conjunction of the human and computational layers in raising awareness in the face of dilemmas.

https://doi.org/10.1590/1982-2553202150301 417 downloads

Characterization and Biological Performance of Marine Sponge Collagen
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Fernandes, Kelly Rossetti

; Parisi, Julia Risso

; Cruz, Matheus de Almeida

; Gabbai-Armelin, Paulo Roberto

; Araújo, Tiago Akira Tashiro de

; Santana, Alan de França

; Avanzi, Ingrid Regina

; Silva-Freitas, Francisca Rayanne

; Ribeiro, Alessandra Mussi

; Vale, Giovanna Caroline Aparecida do

; Fortulan, Carlos Alberto

; Granito, Renata Neves

; Renno, Ana Claudia Muniz

Brazilian Archives of Biology and Technology

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2021, Volume 64 elocation e21200592

Resumo: En | Texto: En | PDF: En

Abstract This study characterized the morphological aspects of marine collagen - spongin (SPG) extract from marine sponges, as well as, evaluating its in vitro and in vivo biological performance. Aplysina fulva marine sponge was used for the SPG extraction. It was investigated the physicochemical and morphological properties of SPG by using scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction and compared to PMMA and bovine collagen. Additionally, the SPG cytotoxicity and its influence on cell proliferation, through in vitro tests. Moreover, the in vivo biological response was investigated using an experimental model of tibial bone defect. The results demonstrated that SPG presented an irregular granular aspect, with a composition of OH, C=O, NH, CN and an amorphous profile. Also, in vitro viability results for the L929 and MC3T3 cell lines cultured with SPG extracts demonstrated normal growth in comparison to controls, except for MC3T3 viability at day 3. For in vivo analysis, using tibial bone defects in rats, SPG treated animals presented an increased rate of material resorption and higher granulation and bone formation deposition in the region of the defect, mainly after 45 days. As a conclusion, SPG was successfully extracted. The in vitro and in vivo studies pointed out that SPG samples produced an increase in L929 and MC3T3 viability and improved the performance in tibial bone defects. It can be concluded that SPG can be used as a bone graft for bone regeneration.

https://doi.org/10.1590/1678-4324-2021200592

Improvement of Mycobacterium tuberculosis detection in sputum using DNA extracted by sonication
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Bello, Graziele Lima

; Morais, Franciele Costa Leite

; Wolf, Jonas Michel

; Gehlen, Mirela

; Soares, Tainá dos Santos

; Halon, Maria Laura

; Barcellos, Regina Bones

; Rossetti, Maria Lucia Rosa

Brazilian Journal of Infectious Diseases

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Out 2020, Volume 24 Nº 5 Páginas 398 - 404

Resumo: En | Texto: En | PDF: En

Abstract Tuberculosis (TB) is one of the infectious diseases with high mortality in the world. DNA amplification techniques have been used to overcome barriers to the diagnosis of this disease. However, the success of these methodologies is highly dependent on the DNA obtained from the sample. This study was carried out to verify whether the DNA extracted by sonication (in house method) could yield suitable DNA for amplification by real-time PCR (qPCR). Sixty sputum samples were submitted to DNA extraction using sonication compared to a commercial method (Detect-TB kit, Labtest/MG-Brazil). All DNA samples were amplified by qPCR for IS6110 region (IS6110-qPCR/SYBR Green assay). Out of 60 samples, 40 were positive for TB; of these, all had positive results when extracted by sonication (100%) and 80% when extracted by the commercial method. The limit of detection (LOD) of Mycobacterium tuberculosis (H37Rv strain) by qPCR was 14CFU/mL when the DNA was extracted by sonication, compared to countless colonies when extracted by commercial kit. In conclusion, the sonication protocol (without purification step) proved to be a simple, fast, and suitable method for obtaining DNA for use in qPCR from sputum samples.

https://doi.org/10.1016/j.bjid.2020.08.006 438 downloads

Rapid detection of Mycobacterium tuberculosis DNA and genetic markers for Isoniazid resistance in Ziehl-Neelsen stained slides
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Bello, Graziele Lima

; Morais, Franciele Costa Leite

; de Jesus, Sheile Pinheiro

; Wolf, Jonas Michel

; Gehlen, Mirela

; de Almeida, Isabela Neves

; Figueiredo, Lida Jouca de Assis

; Soares, Tainá dos Santos

; Barcellos, Regina Bones

; Dalla Costa, Elis Regina

; de Miranda, Silvana Spíndola

; Rossetti, Maria Lucia Rosa

Memórias do Instituto Oswaldo Cruz

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2020, Volume 115 elocation e190407

Resumo: En | Texto: En | PDF: En

BACKGROUND Early diagnosis of tuberculosis (TB) and identification of strains of Mycobacterium tuberculosis resistant to anti-TB drugs are considered the main factors for disease control. OBJECTIVES To standardise a real-time polymerase chain reaction (qPCR) assay technique and apply it to identify mutations involved in M. tuberculosis resistance to Isoniazid (INH) directly in Ziehl-Neelsen (ZN) stained slides. METHODS Were analysed 55 independent DNA samples extracted from clinical isolates of M. tuberculosis by sequencing. For application in TB diagnosis resistance, 59 ZN-stained slides were used. The sensitivity, specificity and Kappa index, with a 95% confidence interval (CI95%), were determined. FINDINGS The agreement between the tests was, for the katG target, the Kappa index of 0.89 (CI95%: 0.7-1.0). The sensitivity and specificity were 97.6% (CI95%: 87.7-99.9) and 91.7% (CI95%: 61.5-99.5), respectively. For inhA, the Kappa index was 0.92 (CI95%: 0.8-1.0), the sensitivity and specificity were 94.4% (CI95%: 72.7-99.8) and 97.3% (CI95%: 85.8-99.9), respectively. The use of ZN-stained slides for drug-resistant TB detection showed significant results when compared to other standard tests for drug resistance. MAIN CONCLUSIONS qPCR genotyping proved to be an efficient method to detect genes that confer M. tuberculosis resistance to INH. Thus, qPCR genotyping may be an alternative instead of sequencing.

https://doi.org/10.1590/0074-02760190407 985 downloads

Factors associated with cognitive performance in elderly caregivers
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Pavarini, Sofia Cristina Iost

; Brigola, Allan Gustavo

; Ottaviani, Ana Carolina

; Luchesi, Bruna Moretti

; Souza, Érica Nestor

; Rossetti, Estefani Serafim

; Moralles, Herick Fernando

; Terassi, Marélli

; Oliveira, Nathalia Alves

; Manzine, Patricia Regina

; Tavares Neto, Roberto Fernandes

Arquivos de Neuro-Psiquiatria

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Out 2018, Volume 76 Nº 10 Páginas 685 - 691

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

RESUMO Objetivos: explorar os fatores socioeconômicos, demográficos e psicossociais associados ao desempenho cognitivo em idosos cuidadores do Brasil. Métodos: Avaliamos 351 idosos cuidadores da atenção primária à saúde em relação a variáveis sociodemográficas e de contexto do cuidado. Os domínios da Escala Cognitiva de Addenbrooke Revisada (ACE-R) - orientação/atenção, memória, fluência verbal, linguagem e visuo-espacial - foram utilizados como variáveis dependentes no modelo de Tobit. Resultados: Alfabetização e renda familiar foram positivamente associados a todos os domínios ACE-R. A idade, o sexo, o tempo de atendimento (dias/semana) foram associados negativamente com alguns domínios cognitivos. Além disso, receber ajuda emocional e nível de esperança foram positivamente associados a domínios específicos. Discussão: os resultados podem ser úteis para o planejamento de intervenções voltadas para cuidadores idosos, a fim de prevenir déficits nos diferentes domínios cognitivos.

Abstract Objectives: To explore the socioeconomic, demographic and psychosocial factors associated with cognitive performance in elderly caregivers from Brazil. Methods: We evaluated 351 Brazilian elderly caregivers attending primary healthcare services regarding sociodemographic and care variables. Addenbrooke's Cognitive Examination-Revised (ACE-R) domains of orientation/attention, memory, verbal fluency, language and visuospatial were used as dependent variables in the Tobit model. Results: Literacy and family income were positively associated with all ACE-R domains. Age, gender, time of care (days/week) were negatively associated with some cognitive domains. Moreover, receiving emotional help and the level of hope were positively associated with specific domains. Discussion: The results may be useful for planning interventions aimed at elderly caregivers in order to prevent deficits in the different cognitive domains.

https://doi.org/10.1590/0004-282x20180101 1468 downloads

A high prevalence of human papillomavirus 16 and 18 co-infections in cervical biopsies from southern Brazil
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Jesus, Sheile Pinheiro de

; Costa, Ana Carla Marques da

; Barcellos, Regina Bones

; Medeiros, Rubia Marília de

; Silva, Cláudia Maria Dornelles da

; Rossetti, Maria Lucia

Brazilian Journal of Microbiology

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2018, Volume 49 Supl. 1 Páginas 220 - 223

Resumo: En | Texto: En | PDF: En

ABSTRACT HPV types 16 and 18 were studied in paraffin-fixed cervical biopsy collected in southern Brazil. HPV 16, HPV 18 and co-infection HPV 16/18 were identified in 10/57 (17.5%), 4/57 (7%) and in 43/57 (75.4%) samples, respectively. Southern Brazil has one of the highest prevalence rates of HPV 16/18 reported.

https://doi.org/10.1016/j.bjm.2018.04.003 879 downloads

Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus
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Santos, Paula Fernanda Gonçalves dos

; Costa, Elis Regina Dalla

; Ramalho, Daniela M

; Rossetti, Maria Lucia

; Barcellos, Regina Bones

; Nunes, Luciana de Souza

; Esteves, Leonardo Souza

; Rodenbusch, Rodrigo

; Anthony, Richard M

; Bergval, Indra

; Sengstake, Sarah

; Viveiros, Miguel

; Kritski, Afrânio

; Oliveira, Martha M

Memórias do Instituto Oswaldo Cruz

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Jun 2017, Volume 112 Nº 6 Páginas 396 - 403

Resumo: En | Texto: En | PDF: En

BACKGROUND To cope with the emergence of multidrug-resistant tuberculosis (MDR-TB), new molecular methods that can routinely be used to screen for a wide range of drug resistance related genetic markers in the Mycobacterium tuberculosis genome are urgently needed. OBJECTIVE To evaluate the performance of multiplex ligaton-dependent probe amplification (MLPA) against Genotype® MTBDRplus to detect resistance to isoniazid (INHr) and rifampicin (RIFr). METHOD 96 culture isolates characterised for identification, drug susceptibility testing (DST) and sequencing of rpoB, katG, and inhA genes were evaluated by the MLPA and Genotype®MTBDRplus assays. RESULTS With sequencing as a reference standard, sensitivity (SE) to detect INHr was 92.8% and 85.7%, and specificity (SP) was 100% and 97.5%, for MLPA and Genotype®MTBDRplus, respectively. In relation to RIFr, SE was 87.5% and 100%, and SP was 100% and 98.8%, respectively. Kappa value was identical between Genotype®MTBDRplus and MLPA compared with the standard DST and sequencing for detection of INHr [0.83 (0.75-0.91)] and RIFr [0.93 (0.88-0.98)]. CONCLUSION Compared to Genotype®MTBDRplus, MLPA showed similar sensitivity to detect INH and RIF resistance. The results obtained by the MLPA and Genotype®MTBDRplus assays indicate that both molecular tests can be used for the rapid detection of drug-resistant TB with high accuracy. MLPA has the added value of providing information on the circulating M. tuberculosis lineages.

https://doi.org/10.1590/0074-02760160376 1493 downloads

10.

Performance of a molecular assay to detect Mycobacterium tuberculosis complex DNA in clinical specimens: multicenter study in Brazil
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Verza, Mirela

; Schmid, Karen Barros

; Barcellos, Regina Bones

; Linck, Natali

; Bello, Graziele Lima

; Scapin, Daniel

; Sperhacke, Rosa Dea

; Silva, Márcia Susana Nunes

; Wollheim, Claudia

; Rivero, Martha Gabriela Celle

; Kritski, Afrânio Lineu

; Rezende Jr, Leonides

; Oliveira, Martha Maria

; Costa, Elis Regina Dalla

; Rossetti, Maria Lucia Rosa

Memórias do Instituto Oswaldo Cruz

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Fev 2017, Volume 112 Nº 2 Páginas 94 - 99

Resumo: En | Texto: En | PDF: En

BACKGROUND In high tuberculosis (TB) burden countries, there are few data on the performance of new molecular commercialised assays developed locally. OBJECTIVE To evaluate the performance of a new molecular commercialised assay for TB diagnosis (Detect-TB) in three laboratories. METHODS A total of 302 sputum samples from an equal number of patients with presumptive diagnosis of pulmonary tuberculosis (PTB) were submitted for routine smear microscopy, culture, and Detect-TB assay at three different sites in Brazil (the cities of Caxias do Sul, São Paulo and Canoas). FINDINGS Seventy four (24.7%) TB cases were diagnosed (65 bacteriologically confirmed). When compared to smear microscopy/culture results, the overall sensitivity and specificity of Detect-TB assay was 84.6% (CI 95%; 73.7-91.6) and 93.1% (CI 95%; 89.1-95.8), respectively. When compared to bacteriological and clinical diagnostic criteria, the sensitivity and specificity of Detect-TB assay was 74.3% (CI 95%; 63.3-82.9) and 92.9% (CI 95%; 88.7-95.6), respectively. Among the three sites - Caxias do Sul, São Paulo and Canoas - the sensitivity and specificity were respectively 94.7% and 97.8%; 71.4% and 93.9%, 82.1% and 88.9%. MAIN CONCLUSIONS These findings suggest that the Detect-TB assay could be applied routinely in reference laboratories across different regions in Brazil.

https://doi.org/10.1590/0074-02760160196 1103 downloads

11.

Low-level laser therapy associated to a resistance training protocol on bone tissue in diabetic rats
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Patrocínio-Silva, Tatiane Lopes

; Souza, André Moreira Fogaça de

; Goulart, Raul Loppi

; Pegorari, Carolina Fuirini

; Oliveira, Jussan Rodrigues

; Fernandes, Kelly Rossetti

; Magri, Angela Maria Paiva

; Pereira, Rosa Maria Rodrigues

; Ribeiro, Daniel Araki

; Nagaoka, Márcia Regina

; Rennó, Ana Claudia Muniz

Archives of Endocrinology and Metabolism

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Out 2016, Volume 60 Nº 5 Páginas 457 - 464

Resumo: En | Texto: En | PDF: En

ABSTRACT Objective The present study aimed to evaluate the in vivo response of a resistance training and low-level laser therapy (LLLT) on tibias and femurs of rats with diabetes mellitus (DM). Materials and methods Forty male Wistar rats were randomly distributed into four experimental groups: control group (CG), diabetic group (DG), diabetic trained group (TG) and diabetic trained and laser irradiated group (TLG). DM was induced by streptozotocin (STZ) and after two weeks laser and resistance training started, performed for 24 sessions, during eight weeks. At the end of the experiment, animals were euthanized and tibias and femurs were removed for analysis. Histological, histomorphometrical, immunohistochemistry and mechanical analyses were performed. Results Trained groups, with or without laser irradiation, showed increased cortical area, bone density and biomechanical properties. The immunohistochemical analysis revealed that TG and TLG demonstrated an increased RUNX2 expression. RANK-L immunoexpression was similar for all experimental groups. Conclusion In conclusion, it can be suggested that the resistance exercise program stimulated bone metabolism, culminating in increased cortical tibial area, bone mineral content, bone mineral density and biomechanical properties. Furthermore, the association of physical exercises and LLLT produced higher values for bone mineral content and stiffness. Consequently, these data highlight the potential of physical exercise in the management of bone loss due to DM and the possible extra osteogenic stimulus offered by lasertherapy. Further long-term studies should be carried out to provide additional information.

https://doi.org/10.1590/2359-3997000000190 1442 downloads

12.

Association of human papillomavirus and Chlamydia trachomatis with intraepithelial alterations in cervix samples
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Wohlmeister, Denise

; Vianna, Débora Renz Barreto

; Helfer, Virgínia Etges

; Gimenes, Fabrícia

; Consolaro, Marcia Edilaine Lopes

; Barcellos, Regina Bones

; Rossetti, Maria Lucia

; Calil, Luciane Noal

; Buffon, Andréia

; Pilger, Diogo André

Memórias do Instituto Oswaldo Cruz

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Fev 2016, Volume 111 Nº 2 Páginas 106 - 113

Resumo: En | Texto: En | PDF: En

The influence of different infectious agents and their association with human papillomavirus (HPV) in cervical carcinogenesis have not been completely elucidated. This study describes the association between cytological changes in cervical epithelium and the detection of the most relevant aetiological agents of sexually transmitted diseases. Samples collected from 169 patients were evaluated by conventional cytology followed by molecular analysis to detect HPV DNA, Chlamydia trachomatis, herpes simplex virus 1 and 2,Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, andTreponema pallidum, besides genotyping for most common high-risk HPV. An association between cytological lesions and different behavioural habits such as smoking and sedentariness was observed. Intraepithelial lesions were also associated with HPV and C. trachomatis detection. An association was also found between both simple and multiple genotype infection and cytological changes. The investigation of HPV and C. trachomatisproved its importance and may be considered in the future for including in screening programs, since these factors are linked to the early diagnosis of patients with precursor lesions of cervical cancer.

https://doi.org/10.1590/0074-02760150330 3275 downloads

13.

Association between cervical lesion grade and micronucleus frequency in the Papanicolaou test
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Bueno, Caroline Tanski

; Silva, Cláudia Maria Dornelles da

; Barcellos, Regina Bones

; Silva, Juliana da

; Santos, Carla Rossana dos

; Menezes, João Evangelista Sampaio

; Menezes, Honório Sampaio

; Rossetti, Maria Lucia Rosa

Genetics and Molecular Biology

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Set 2014, Volume 37 Nº 3 Páginas 496 - 499

Resumo: En | Texto: En | PDF: En

The aim of this study was to evaluate the association between the frequency of micronuclei (MN) and the cellular changes detected in the conventional Papanicolaou test. One hundred and seventy-four Papanicolaou test smears with cellular changes were examined. MN screening was done in cytopathological smears by counting 1,000 cervical cells in a light microscope. MN frequencies were significantly higher in the group with cellular changes compared to the control group (p < 0.001). The mean MN frequencies were 0.95 ± 1.12 (mean ± SD) in the control group (n = 223), 2.98 ± 1.20 in individuals with atypical squamous cells of undetermined significance (ASC-US) (n = 50), 4.04 ± 1.45 in cervical intraepithelial neoplasia (CIN) I (n = 52), 5.97 ± 1.83 in CIN II (n = 30), 7.29 ± 1.55 in CIN III (n = 17) and 8.64 ± 1.55 in invasive cancer (n = 25). These findings suggest that MN monitoring should be included as an additional criterion for the early detection of cytogenetic damage in routine examinations. This monitoring should be done in the same smear as used for cytopathological examination. More specific and systematic studies are necessary to confirm this proposal.

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14.

Association between human papillomavirus (HPV) DNA and micronuclei in normal cervical cytology
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Cassel, Ana Paula Rebelo

; Barcellos, Regina Bones

; Silva, Cláudia Maria Dornelles da

; Almeida, Sabrina Esteves de Matos

; Rossetti, Maria Lucia Rosa

Genetics and Molecular Biology

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Jun 2014, Volume 37 Nº 2 Páginas 360 - 363

Resumo: En | Texto: En | PDF: En

The aim of the present study was to investigate the association between HPV-DNA and micronucleus (MN) frequency in women with normal cervical cytology. A total of 158 normal cervical smears were analyzed cytologically. The HPV genome was amplified using the GP5+/bioGP6+ consensus primers. HPV-DNA of high-risk types 16, 18, 31, 33, 39, 45 and 59 were also investigated. Of the 158 samples, 20 (12.7%) and 47 (29.7%) were positive for HPV-DNA and MN, respectively. Evidence for MN was found in 11 out of 20 (55%) HPV-DNA positive samples and in 36 out of 138 (26.1%) HPV-DNA negative ones. MN presence was significantly higher in HPV-DNA positive samples (p = 0.016). On the other hand, the absence of MN observed in a considerable number of HPV-DNA negative samples (102) may be of great value in predicting the absence of HPV. The mean age of HPV-DNA positive women (34.2 ± 12.6) was significantly lower than the mean age of HPV-DNA negative women (43.9 ± 13.7) (p = 0.003). Infection by one or multiple HPV types was found in 11 out of 20 (55.0%) and 9 out of 20 (45.0%) samples, respectively. The evaluation of MN using cervical smears collected for cytology tests could, thus, be used as additional information to monitor a population's exposure to HPV.

1543 downloads

15.

In house reverse membrane hybridisation assay versus GenoType MTBDRplus and their performance to detect mutations in the genes rpoB, katG and inhA
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Ferreira Junior, Sergio Luiz Montego

; Costa, Elis Regina Dalla

; Santos, Paula Gonçalves dos

; Gomes, Harrison Magdinier

; Silva, Marcia Susana Nunes

; Esteves, Leonardo Souza

; Oliveira, Martha Maria

; Maschmann, Raquel de Abreu

; Kritski, Afrânio Lineu

; Suffys, Philip Noel

; Rossetti, Maria Lucia Rosa

Memórias do Instituto Oswaldo Cruz

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Jun 2014, Volume 109 Nº 3 Páginas 307 - 314

Resumo: En | Texto: En | PDF: En

Drug-resistant tuberculosis (TB) threatens global TB control and is a major public health concern in several countries. We therefore developed a multiplex assay (LINE-TB/MDR) that is able to identify the most frequent mutations related to rifampicin (RMP) and isoniazid (INH) resistance. The assay is based on multiplex polymerase chain reaction, membrane hybridisation and colorimetric detection targeting of rpoB and katG genes, as well as the inhA promoter, which are all known to carry specific mutations associated with multidrug-resistant TB (MDR-TB). The assay was validated on a reference panel of 108 M. tuberculosis isolates that were characterised by the proportion method and by DNA sequencing of the targets. When comparing the performance of LINE-TB/MDR with DNA sequencing, the sensitivity, specificity and agreement were 100%, 100% and 100%, respectively, for RMP and 77.6%, 90.6% and 88.9%, respectively, for INH. Using drug sensibility testing as a reference standard, the performance of LINE-TB/MDR regarding sensitivity, specificity and agreement was 100%, 100% and 100% (95%), respectively, for RMP and 77%, 100% and 88.7% (82.2-95.1), respectively, for INH. LINE-TB/MDR was compared with GenoType MTBDRplus for 65 isolates, resulting in an agreement of 93.6% (86.7-97.5) for RIF and 87.4% (84.3-96.2) for INH. LINE-TB/MDR warrants further clinical validation and may be an affordable alternative for MDR-TB diagnosis.

https://doi.org/10.1590/0074-0276130469 1524 downloads

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