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Influenza A(H3N2) infection followed by separate COVID-19 infection
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Gregianini, Tatiana Schäffer

; Salvato, Richard Steiner

; Baethgen, Ludmila Fiorenzano

; Piazza, Cláudia Fasolo

; Barcellos, Regina Bones

; Godinho, Fernanda Marques

; Veiga, Ana Beatriz Gorini da

Revista Panamericana de Salud Pública

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2023, Volume 47 elocation e61

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

RESUMEN Este estudio describe el caso de un profesional de la salud que contrajo la infección primero por el virus de la gripe A (H3N2) y a continuación por el coronavirus 2 del síndrome respiratorio agudo grave (SARS-CoV-2) 11 días después. Se recogieron muestras respiratorias y datos clínicos del paciente y sus contactos cercanos. Se extrajo ARN de muestras y se utilizó la reacción en cadena de la polimerasa cuantitativa con transcripción inversa (RT-qPCR, por su sigla en inglés) para investigar los virus. El paciente presentó dos procesos infecciosos distintos: el primero se caracterizó por fiebre, dolor corporal y torácico, postración y cansancio, que cesó en el noveno día. La prueba mediante RT-qPCR solo fue positiva en el virus de la gripe A (H3N2). Once días después del inicio de los primeros síntomas, el paciente manifestó dolor de garganta, congestión nasal, catarro, picazón nasal, estornudos y tos. Una segunda prueba mediante RT-qPCR solo fue positiva para el SARS-CoV-2 y durante este segundo proceso los síntomas duraron 11 días. La secuenciación del SARS-CoV-2 identificó el linaje ómicron BA.1. De los contactos del paciente, uno presentaba una coinfección por el virus de la gripe A (H3N2) y el linaje BA.1.15 del SARS-COV-2, y los otros dos presentaban infecciones únicamente por SARS-CoV-2, uno también del linaje ómicron BA.1.15 y el otro de BA.1.1. Estos hallazgos refuerzan la importancia de realizar pruebas para detectar diferentes virus en casos de sospecha de infección viral respiratoria durante la vigilancia epidemiológica de rutina porque las manifestaciones clínicas comunes de COVID-19 son similares a las de otros virus, como en el caso de la gripe.

RESUMO Este estudo descreve o caso de uma profissional de saúde infectada primeiro pelo vírus influenza A (H3N2) e, 11 dias depois, pelo coronavírus da síndrome respiratória aguda grave 2 (SARS-CoV-2). Amostras respiratórias e dados clínicos foram coletados da paciente e de contatos próximos. RNA foi extraído das amostras, e o método de reação em cadeia da polimerase via transcriptase reversa quantitativa (RT-qPCR) foi utilizado para investigar os vírus. A paciente apresentou dois quadros clínicos distintos. O primeiro foi caracterizado por febre, dor no peito e no corpo, prostração e fadiga, que cessou no nono dia. A RT-qPCR foi positiva apenas para o vírus da influenza A (H3N2). Onze dias após o início dos primeiros sintomas, a paciente apresentou dor de garganta, congestão nasal, coriza, prurido nasal, espirros e tosse. Um segundo teste de RT-qPCR foi positivo apenas para SARS-CoV-2. No segundo evento, os sintomas duraram 11 dias. O sequenciamento do SARS-CoV-2 identificou a cepa Ômicron BA.1. Dentre os contatos da paciente, um teve coinfeção por influenza A (H3N2) e SARS-COV-2 (cepa BA.1.15), e os outros dois foram infectados apenas por SARS-CoV-2 (um também pela cepa Ômicron BA.1.15 e o outro pela BA.1.1). Nossos achados reforçam a importância de testes para a detecção de diferentes vírus em casos de suspeita de infecção viral respiratória durante a vigilância epidemiológica de rotina, visto que as manifestações clínicas comuns da COVID-19 imitam as de outros vírus, como o vírus influenza.

ABSTRACT This study describes the case of a health professional infected first by influenza virus A(H3N2) and then by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 11 days later. Respiratory samples and clinical data were collected from the patient and from close contacts. RNA was extracted from samples and reverse transcription–quantitative polymerase chain reaction (RT-qPCR) was used to investigate the viruses. The patient presented with two different illness events: the first was characterized by fever, chest and body pain, prostration and tiredness, which ceased on the ninth day; RT-qPCR was positive only for influenza virus A(H3N2). Eleven days after onset of the first symptoms, the patient presented with sore throat, nasal congestion, coryza, nasal itching, sneezing and coughing, and a second RT-qPCR test was positive only for SARS-CoV-2; in the second event, symptoms lasted for 11 days. SARS-CoV-2 sequencing identified the Omicron BA.1 lineage. Of the patient’s contacts, one was coinfected with influenza A(H3N2) and SARS-CoV-2 lineage BA.1.15 and the other two were infected only with SARS-CoV-2, one also with Omicron BA.1.15 and the other with BA.1.1. Our findings reinforce the importance of testing for different viruses in cases of suspected respiratory viral infection during routine epidemiological surveillance because common clinical manifestations of COVID-19 mimic those of other viruses, such as influenza.

https://doi.org/10.26633/rpsp.2023.61

Chikungunya virus infection in the southernmost state of Brazil was characterised by self-limited transmission (2017-2019) and a larger 2021 outbreak
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Gregianini, Tatiana Schäffer

; Salvato, Richard Steiner

; Barcellos, Regina Bones

; Godinho, Fernanda Marques

; Ruivo, Amanda Pellenz

; de Melo, Viviane Horn

; Schroder, Júlio Augusto

; Martiny, Fernanda Letícia

; Möllmann, Erica Bortoli

; Favreto, Cátia

; Baethgen, Ludmila Fiorenzano

; Ferreira, Vithoria Pompermaier

; de Lima, Lívia Eidt

; Piazza, Cláudia Fasolo

; Machado, Taís Raquel Marcon

; Becker, Irina Marieta

; Ramos, Raquel Rocha

; Frölich, Guilherme Carey

; Rossetti, Alana Fraga

; Almeida, Lucas da Cunha

; Rodrigues, Tahiana Machado Antunes

; Bragança, Isabella Tabelli

; Campos, Aline Alves Scarpellini

; Manzoni, Verônica Baú

; Machado, Lais Ceschini

; da Silva, Luisa Maria Inácio

; de Oliveira, André Luiz Sá

; Paiva, Marcelo Henrique Santos

; Nunes, Zenaida Marion Alves

; de Almeida, Paula Rodrigues

; Demoliner, Meriane

; Gularte, Juliana Schons

; da Silva, Mariana Soares

; Filippi, Micheli

; Pereira, Vyctoria Malayhka de Abreu Góes

; Spilki, Fernando Rosado

; da Veiga, Ana Beatriz Gorini

; Wallau, Gabriel Luz

Memórias do Instituto Oswaldo Cruz

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2023, Volume 118 elocation e220259

Abstract: En | Text: En | PDF: En

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.

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

Abstract: En | Text: 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

Molecular epidemiology of Mycobacterium tuberculosis in Brazil before the whole genome sequencing era: a literature review
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Conceição, Emilyn Costa

; Salvato, Richard Steiner

; Gomes, Karen Machado

; Guimarães, Arthur Emil dos Santos

; da Conceição, Marília Lima

; Souza e Guimarães, Ricardo José de Paula

; Sharma, Abhinav

; Furlaneto, Ismari Perini

; Barcellos, Regina Bones

; Bollela, Valdes Roberto

; Anselmo, Lívia Maria Pala

; Sisco, Maria Carolina

; Niero, Cristina Viana

; Ferrazoli, Lucilaine

; Refrégier, Guislaine

; Lourenço, Maria Cristina da Silva

; Gomes, Harrison Magdinier

; de Brito, Artemir Coelho

; Catanho, Marcos

; Duarte, Rafael Silva

; Suffys, Philip Noel

; Lima, Karla Valéria Batista

Memórias do Instituto Oswaldo Cruz

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2021, Volume 116 elocation e200517

Abstract: En | Text: En | PDF: En

Molecular-typing can help in unraveling epidemiological scenarios and improvement for disease control strategies. A literature review of Mycobacterium tuberculosis transmission in Brazil through genotyping on 56 studies published from 1996-2019 was performed. The clustering rate for mycobacterial interspersed repetitive units - variable tandem repeats (MIRU-VNTR) of 1,613 isolates were: 73%, 33% and 28% based on 12, 15 and 24-loci, respectively; while for RFLP-IS6110 were: 84% among prison population in Rio de Janeiro, 69% among multidrug-resistant isolates in Rio Grande do Sul, and 56.2% in general population in São Paulo. These findings could improve tuberculosis (TB) surveillance and set up a solid basis to build a database of Mycobacterium genomes.

https://doi.org/10.1590/0074-02760200517 744 downloads

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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oct 2020, Volume 24 N. 5 Pages 398 - 404

Abstract: En | Text: 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

Abstract: En | Text: 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

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 Suppl. 1 Pages 220 - 223

Abstract: En | Text: 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 N. 6 Pages 396 - 403

Abstract: En | Text: 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

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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feb 2017, Volume 112 N. 2 Pages 94 - 99

Abstract: En | Text: 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

10.

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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feb 2016, Volume 111 N. 2 Pages 106 - 113

Abstract: En | Text: 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

11.

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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sep 2014, Volume 37 N. 3 Pages 496 - 499

Abstract: En | Text: 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.

1569 downloads

12.

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 N. 2 Pages 360 - 363

Abstract: En | Text: 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.

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