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1.
Rev. patol. trop ; 49(1)2020. tab
Article in English | LILACS | ID: biblio-1099655

ABSTRACT

Colonial cheese is a culturally and economically important product from the south of Brazil. As most of its production is artisanal, the technology employed is mostly knowledge passed down from one generation to the next according to family tradition and may be produced with raw or pasteurized milk. It is noted for its spicy flavour and variable composition and is often classified as a medium to high-moisture cheese. This intrinsic feature increases the risk of microbial spoilage and food poisoning. One of the main bio-indicators of contamination in colonial cheese is coagulase positive Staphylococcus. The purpose of this study was the phenotypic identification of Staphylococcus species isolated from the products and surfaces in the main production stages of colonial cheese. Staphylococcus sp. isolates from the food and the production environment were obtained from two colonial cheese-production agro-industries in Rio Grande do Sul. Samples of fresh milk, curd, ripening and final colonial cheese were collected. In addition, surface sampling was performed on the coagulation tanks, production tables, molds, cheese ripening shelves and on the hands of the handlers. Staphylococcus sp. isolates in the cheese and the production environments tested in this study were identified by phenotypic techniques through biochemical and MALDI-TOF MS analyses. These isolates were subjected to gene expression analysis for enterotoxins A, B, C, D, and E. All isolates (72) were identified as Staphylococcus sp., and 43% of the total isolates tested were coagulase positive. Staphylococcus aureus was the predominant species in the raw milk and production tanks. Regarding coagulase negative staphylococci isolates, S. warneri and S. sciuri were most abundant. The sea and seb genes were detected in 4% of the Staphylococcus isolates. The results indicate eleven different species of Staphylococcus present in the colonial cheese production environments studied. The predominant presence of S. aureus in the different samples of milk, curd, ripened cheese, ready-to-eat cheese and hands of the handlers indicates that there are issues with the selection of milk-producing animals, pasteurization process and/or hygiene control of handlers. The sea and seb genes were detected in samples of raw milk and colonial cheese. No enterotoxin genes were detected in coagulase negative staphylococci.


Subject(s)
Staphylococcus , Cheese/analysis , Coagulase , Enterotoxins
2.
Clin. biomed. res ; 39(4): 322-332, 2019.
Article in English | LILACS | ID: biblio-1087323

ABSTRACT

O transtorno por uso de álcool (TUA) é influenciado pela genética, principalmente na metabolização do etanol. Os genes da álcool desidrogenase (ADH1B/ADH1C), enzima que transforma o etanol, apresentam SNPs (single nucleotide polymorphisms) que resultam em isoenzimas com diferentes taxas catalíticas. Estudos demonstraram que os SNPs Arg48His, Arg370Cys, Arg272Gln e Ile350Val contribuem para o TUA. Este artigo revisou os estudos que investigaram SNPs em ADH1B (Arg48His/Arg370Cys) e ADH1C (Arg272Gln/Ile350Val), bem como avaliou as variações nas frequências alélicas desses genes e a influência no TUA nas diferentes populações no mundo. As frequências alélicas dos polimorfismos foram comparadas pelos testes qui-quadrado de Pearson e exato de Fisher (p < 0,05). O SNP Arg48His confere proteção para o TUA em euroamericanos, latino-americanos, europeus, brasileiros, asiáticos e australianos. O SNP Arg370Cys confere proteção para o TUA em afrodescendentes. Os SNPs Arg272Gln e Ile350Val predispõem o TUA principalmente em europeus. Os SNPs Arg48His, Arg370Cys e Arg272Gln/Ile350Val foram mais frequentes em amostras de leste-asiáticos (69,7%), africanos (19,1%) e europeus (40,5%), respectivamente (p < 0,01). Os diferentes alelos dos genes ADH1B/ADH1C devido a SNPs têm uma importante contribuição no TUA. As frequências desses alelos variam conforme a população, resultando em diferentes efeitos no TUA. (AU)


Alcohol use disorder (AUD) is influenced by genetics, especially in the metabolism of ethanol. The ethanol dehydrogenase genes (ADH1B/ADH1C), which convert ethanol, have single nucleotide polymorphisms (SNPs) that result in isoenzymes with different catalytic rates. Studies have shown that the Arg48His, Arg370Cys, Arg272Gln, and Ile350Val SNPs contribute to AUD. This article reviewed the studies that investigated SNPs in ADH1B (Arg48His/Arg370Cys) and ADH1C (Arg272Gln/Ile350Val) and evaluated variations in the allele frequencies of these genes and their influence on AUD in different populations worldwide. The allele frequencies of the polymorphisms were compared by Pearson's chi-square and Fisher's exact tests (p < 0.05). The Arg48His SNP provides protection against AUD in Euro-Americans, Latin Americans, Europeans, Brazilians, Asians, and Australians. The Arg370Cys SNP provides protection against AUD in Afro-descendants. The Arg272Gln and Ile350Val SNPs predispose to AUD mainly in Europeans. The Arg48His, Arg370Cys, and Arg272Gln/Ile350Val SNPs were more frequent in East Asians (69.7%), Africans (19.1%), and Europeans (40.5%), respectively (p < 0.01). The different alleles of the ADH1B/ADH1C genes due to SNPs make an important contribution to AUD. The frequencies of these alleles vary among different populations, resulting in different effects on AUD..(AU)


Subject(s)
Humans , Alcohol-Related Disorders/genetics , Polymorphism, Single Nucleotide/genetics , Alcohol Dehydrogenase/biosynthesis , Alcohol-Related Disorders/epidemiology , Ethanol/adverse effects
3.
Braz. j. infect. dis ; 22(5): 424-432, Sept.-Oct. 2018. tab, graf
Article in English | LILACS | ID: biblio-974235

ABSTRACT

ABSTRACT Introduction: Nontyphoidal Salmonella serotypes are the main cause of human food-borne infection, including several hospitalization cases in the developing countries. Aim: To detect the main serotypes and to characterize the antibiotic resistance of human non-enteric and enteric nontyphoidal Salmonella from clinical isolates in Brazil. Methods: Salmonella serotypes were identified by microbiological and molecular methods. Susceptibility testing to antibiotics was performed by agar disk diffusion. Real-time PCRs were carried out for the detection of the genus Salmonella as well as serotypes Typhimurium and Enteritidis. Results: A total of 307 nontyphoidal Salmonella were isolated from 289 different patients in a reference laboratory (LACEN-RS) from Southern Brazil in a six-year period (2010-2015). There were 45 isolates from emerging cases and 244 from sporadic cases in hospitalized patients. Non-enteric isolates were detected in 42.6% of the patients from sources such as urine, blood and other clinical fluids. Serological and PCR-specific tests demonstrated that Typhimurium (48.4%) and Enteritidis (18.3%) were the most frequent serotypes. Typhimurium isolates were generally resistant to three or more antibiotic classes, while Enteritidis isolates to one or two classes. Typhimurium was the most frequent serotype in all samples (48.4%), mainly among the hospitalized patients (55.6%), and presented the highest rates of multidrug resistance (59.3% of the isolates of this serotype). Further, the prevalence of this serotype increased along the years of the study in comparison to other nontyphoidal Salmonella serotypes. Conclusion: Greater public health attention should be given to prevent salmonellosis in the community and in hospital settings to reduce the rates of Typhimurium strains with multidrug resistance.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Young Adult , Salmonella Infections/microbiology , Salmonella Infections/epidemiology , Salmonella typhimurium/drug effects , Drug Resistance, Multiple, Bacterial , Hospitalization/statistics & numerical data , Salmonella enteritidis/isolation & purification , Salmonella enteritidis/drug effects , Salmonella typhimurium/isolation & purification , Time Factors , Brazil/epidemiology , Microbial Sensitivity Tests , Serotyping , Cross Infection/microbiology , Cross Infection/epidemiology , Prevalence , Real-Time Polymerase Chain Reaction , Serogroup , Anti-Bacterial Agents/pharmacology
4.
Braz. j. infect. dis ; 22(4): 294-304, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-974222

ABSTRACT

ABSTRACT Background Hepatitis B virus (HBV) infection is a major public health problem in Brazil. HBV endemicity is usually moderate to low according to geographic regions, and high prevalence of this virus has been reported in people of some specific Brazilian counties, including those with a strong influence of Italian colonization in southern Brazil. Analysis of HBV diversity and identification of the main risk factors to HBV infection are necessary to understand hepatitis B epidemiology in these high prevalence regions in southern Brazil. Objective To investigate epidemiological characteristics and HBV genotypes and subgenotypes circulating in a specific city with high HBV prevalence. Methods A cross-sectional study was performed with 102 HBV chronically infected individuals, recruited in reference outpatient clinics for viral hepatitis in a city of high HBV prevalence (Bento Gonçalves) in Rio Grande do Sul state, Brazil between July and December 2010. Socio-demographic, clinical and behavior-related variables were collected in a structured questionnaire. HBV serological markers (HBsAg, anti-HBc), viral load, genotypes/subgenotypes and drug resistance were evaluated and comparatively analyzed among all patients. Results The HBV infected subjects had a mean age of 44.9 (±12.2) years, with 86 patients (84.3%) reporting to have a family history of HBV infection, 51 (50.0%) to share personal objects, and were predominantly of Italian descendants (61; 64.9%). There was a predominance of genotype D (49/54; 90.7%), but genotype A was also detected (5/54; 9.3%). Subgenotypes D1 (1; 4.7%), D2 (3; 14.3%), and D3 (17; 81.0%) were identified. LAM-resistant mutation (rtM204I) and ADV-resistant mutations (rtA181V) were detected in only one patient each. Conclusions These results demonstrate a pivotal role of intrafamilial transmission for HBV spreading in this population. Furthermore, there is a high prevalence of HBV genotype D in this region.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Young Adult , Hepatitis B virus/genetics , Hepatitis B, Chronic/epidemiology , Drug Resistance, Viral , Antiviral Agents/therapeutic use , Brazil/epidemiology , Hepatitis B virus/drug effects , Polymerase Chain Reaction , Prevalence , Cross-Sectional Studies , Risk Factors , Viral Load , Hepatitis B, Chronic/virology , Genotype , Hepatitis B Surface Antigens/blood , Mutation
5.
Genet. mol. biol ; 41(1): 92-97, Jan.-Mar. 2018. tab
Article in English | LILACS-Express | LILACS | ID: biblio-892476

ABSTRACT

Abstract Functional dyspepsia and lactose intolerance (adult-type hypolactasia, ATH) are common conditions that may coexist or even be confounded. Their clinical presentation can be similar, however, lactose intolerance does not form part of the diagnostic investigation of functional dyspepsia. Studies on the association between functional dyspepsia and ATH are scarce. This study aimed to evaluate whether ATH is associated with symptoms of functional dyspepsia. Patients fulfilling the Rome III diagnostic criteria for functional dyspepsia underwent genetic testing for ATH. Dyspeptic symptoms were evaluated and scored according to a validated questionnaire. The diagnostic criteria for ATH was a CC genotype for the -13910C/T polymorphism, located upstream of the lactase gene. The mean scores for dyspeptic symptoms were compared between patients with ATH and those with lactase persistence. A total of 197 functional dyspeptic patients were included in the study. Mean age was 47.7 years and 82.7% patients were women. Eighty-eight patients (44.7%) had a diagnosis of ATH. Abdominal bloating scores were higher in ATH patients compared to the lactase persistent patients (P=0.014). The remaining dyspeptic symptom scores were not significantly different between the two groups. The study results demonstrate an association between ATH and bloating in patients with functional dyspepsia.

6.
Rev. Soc. Bras. Med. Trop ; 51(1): 30-38, Jan.-Feb. 2018. tab, graf
Article in English | LILACS, ColecionaSUS, CONASS, SES-RS | ID: biblio-897050

ABSTRACT

INTRODUCTION Infections caused by respiratory viruses are important problems worldwide, especially in children. Human metapneumovirus (hMPV) is a respiratory pathogen and causes severe infections with nonspecific symptoms. This study reports the hMPV occurrence and dissemination in southern Brazil and compares the frequency of occurrence of this virus and the human respiratory syncytial virus (hRSV) in the epidemiological weeks in a three-year period (2009-2011). METHODS: In total, 545 nasopharyngeal (NP) specimens from individuals with Severe Acute Respiratory Syndrome (SARS) who were negative for other seven respiratory viruses were analyzed for the presence of hMPV. Human metapneumovirus was detected by direct immunofluorescence and real-time reverse transcription polymerase chain reaction. RESULTS: hMPV was detected in 109 patients from the main geographic regions of the southernmost state of Brazil, presenting similar overall prevalence in males (46.8%) and females (53.2%). Among children who were less than six years old, hMPV was detected in 99 samples of all age groups, with a higher frequency in infants who were less than one year old (45.7%) compared to all other age groups until six years. hMPV and hRSV infection occurred in almost the same epidemiological weeks (EWs) of each year, with peaks of incidence between EW 31/37 and EW 26/38 for the years 2009 and 2011, respectively. hMPV was further detected in several cases of SARS and it was the only virus detected in three deaths. CONCLUSIONS These findings indicate that hMPV is in circulation in southern Brazil and highlight the importance of diagnosing hMPV for influenza-like illness in the population. (AU)


Subject(s)
Humans , Male , Female , Pregnancy , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Respiratory Tract Infections/transmission , Respiratory Tract Infections/virology , Respiratory Syncytial Virus Infections/virology , Metapneumovirus/pathogenicity , Epidemiological Monitoring , Adenoviruses, Human , Pneumovirinae/classification , Paramyxoviridae Infections/virology , Coronavirus , Enterovirus , Severe Acute Respiratory Syndrome , Influenza, Human , Human bocavirus
7.
Mem. Inst. Oswaldo Cruz ; 112(8): 544-550, Aug. 2017. tab
Article in English | LILACS | ID: biblio-894870

ABSTRACT

BACKGROUND Hepatitis B virus (HBV) infection is a major public health problem in Brazil. Several risk factors are involved in HBV infection and their identification by a rational and essential approach is required to prevent the transmission of this infection in Brazil. OBJECTIVES To evaluate risk factors associated with HBV infection in South Brazil. METHODS A total of 260 patients with HBV and 260 controls from Caxias do Sul (state of Rio Grande do Sul, Brazil) participated in this study. All participants were given a standard questionnaire to yield the sociodemographic information and to identify HBV risk factors. HBV infection was detected by HBsAg test in all participants. FINDINGS HBV infection in these cases was strongly associated with history of a family member HBV-infected, mainly mother [odds ratio (OR) = 4.86; 95% confidence intervals (CI): 1.69-13.91], father (OR = 5.28; 95% CI: 1.58-17.71), and/or siblings (OR = 22.16; 95% CI: 9.39-52.25); sharing personal objects (OR = 1.40; 95% CI: 1.37-2.38); and having history of blood transfusion (OR = 2.05; 95% CI: 1.10-2.84). CONCLUSIONS HBV infection was strongly associated with having a family member infected with hepatitis B, sharing personal objects, and having history of blood transfusion.


Subject(s)
Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Hepatitis B virus/immunology , Hepatitis B, Chronic/diagnosis , Hepatitis B, Chronic/transmission , Hepatitis B, Chronic/epidemiology , Hepatitis B Antibodies/blood , Hepatitis B Surface Antigens/blood , Socioeconomic Factors , Brazil/epidemiology , Case-Control Studies , Family Health , Transfusion Reaction
8.
Clin. biomed. res ; 37(3): 214-231, 2017. tab, ilus
Article in Portuguese | LILACS | ID: biblio-859835

ABSTRACT

A ingestão de bebidas alcoólicas é um evento socioculturalmente aceito em muitos países. Porém, o consumo frequente e descontrolado deste tipo de bebida configura o transtorno por uso de álcool (TUA). Esta condição causa agravos que podem afetar a sociedade de uma forma geral. O TUA também pode levar os pacientes a contraírem doenças. Entre estas, existe uma relação importante entre TUA e doenças infectocontagiosas, com destaque para a infecção pelo HIV e o posterior desenvolvimento da AIDS. Portanto, a presente pesquisa objetivou realizar uma revisão da literatura sobre as relações entre TUA e HIV/AIDS. A seleção do material científico foi efetuada tendo por base plataformas eletrônicas, tais como: Google Scholar, MEDLINE, LILACS, SciELO, NCBI / PUBMED, Scopus e Science Direct. O entendimento dos fatores relacionados ao TUA, principalmente em pacientes com HIV/AIDS, é de fundamental importância para a formulação e criação de estratégias de políticas públicas que visem reduzir esta possível relação (AU)


The ingestion of alcoholic beverages is socio-culturally accepted in many countries. However, frequent and uncontrolled consumption of this type of beverage constitutes alcohol use disorder (AUD). This condition may be harmful to society in general, and it can lead patients to contract other diseases. There is an important relationship between AUD and infectious diseases, with emphasis on HIV infection and the later development of AIDS. Therefore, the present research aimed to carry out a review of the literature on the relationship between AUD and HIV/AIDS. The selection of the scientific material was based on electronic platforms, such as Google Scholar, MEDLINE, LILACS, SciELO, NCBI/ PUBMED, Scopus and Science Direct. The understanding of the factors related to AUD, especially in patients with HIV/AIDS, is of fundamental importance for the formulation and creation of public policy strategies aimed at reducing this possible relationship (AU)


Subject(s)
Humans , Alcohol-Related Disorders/complications , HIV Infections/transmission , Alcohol Drinking/adverse effects , HIV Infections/chemically induced , Viral Load/physiology , Virus Replication/physiology
9.
Braz. j. infect. dis ; 20(1): 61-68, Jan.-Feb. 2016. tab
Article in English | LILACS | ID: lil-776455

ABSTRACT

Abstract Human papillomavirus (HPV) infection is common in sexually active women and viral persistence may cause intraepithelial lesions and eventually progress to cervical cancer (CC). The present study aimed to investigate epidemiological factors related to HPV infection and to evaluate viral persistence and CC precursor lesions frequencies in women from a city in the countryside of South Brazil. Three hundred women were recruited from a primary public health care clinic. The patients were interviewed and underwent sampling with cervical brushes for HPV-DNA detection/typing by a PCR-based assay and cytological analysis by Pap smear test. HPV was detected in 47 (15.7%) women. HPV infection was significantly associated with young age (<30 years) and low socio-economic status. Seventeen (5.7%) women presented cytological abnormalities, three of them with precursor CC intraepithelial lesions. A subgroup of 79 women had been previously analyzed and thirteen (16.4%) were persistently infected, two with precursor CC intraepithelial lesions and high-risk HPV types infection (both of them without cervical abnormalities in the first exam). In conclusion, HPV infection was associated with young age (<30 years) and low family income; viral persistence was low (16.4%) but related to CC precursor lesions; and HPV-DNA high risk types detection would help to screen CC in the population.


Subject(s)
Adult , Female , Humans , Middle Aged , Young Adult , Papillomavirus Infections/diagnosis , Uterine Cervical Neoplasms/virology , Brazil/epidemiology , Cross-Sectional Studies , DNA, Viral/analysis , Papanicolaou Test , Prevalence , Papillomaviridae/genetics , Papillomavirus Infections/complications , Papillomavirus Infections/epidemiology , Risk Factors , Socioeconomic Factors , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/epidemiology , Vaginal Smears
10.
Rev. Soc. Bras. Med. Trop ; 48(3): 249-257, May-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-749868

ABSTRACT

INTRODUCTION: Human immunodeficiency virus type 1 (HIV-1) has spread worldwide, with several subtypes and circulating recombinant forms. Brazil has an incidence of 20.5 HIV-1/acquired immunodeficiency syndrome (AIDS) patients per 100,000 inhabitants; however, the Southernmost State of Rio Grande do Sul (RS) has more than twice the number of HIV-1-infected people (41.3/100,000 inhabitants) and a different pattern of subtype frequencies, as previously reported in studies conducted in the capital (Porto Alegre) and its metropolitan region. This study examined HIV-1/AIDS epidemiological and molecular aspects in the countryside of Rio Grande do Sul. METHODS: Socio-demographic, clinical and risk behavioral characteristics were obtained from HIV-1-positive adult patients using a structured questionnaire. HIV-1 subtypes were determined by nested-polymerase chain reaction (PCR) and sequencing of the pol and env genes. RESULTS: The study sample included 149 (55% women) patients with a mean age of 41.8 ± 11.9 years. Most (73.8%) patients had a low education level and reported heterosexual practices as the most (91.9%) probable transmission route. HIV-1 subtypes were detected in 26 patients: 18 (69.2%) infected with subtype C, six (23.1%) infected with subtype B and two (7.7%) infected with BC recombinant forms. CONCLUSIONS: These data highlight the increasing number of HIV-1 subtype C infections in the countryside of South Brazil. .


Subject(s)
Adult , Female , Humans , Male , HIV Infections/epidemiology , HIV-1 , Brazil/epidemiology , Cross-Sectional Studies , Genotype , Genes, env/genetics , Genes, gag/genetics , HIV Infections/virology , HIV-1 , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Risk Factors
12.
Rev. Soc. Bras. Med. Trop ; 47(3): 287-294, May-Jun/2014. tab, graf
Article in English | LILACS | ID: lil-716399

ABSTRACT

Introduction Molecular biology procedures to detect, genotype and quantify hepatitis C virus (HCV) RNA in clinical samples have been extensively described. Routine commercial methods for each specific purpose (detection, quantification and genotyping) are also available, all of which are typically based on polymerase chain reaction (PCR) targeting the HCV 5′ untranslated region (5′UTR). This study was performed to develop and validate a complete serial laboratory assay that combines real-time nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) techniques for the complete molecular analysis of HCV (detection, genotyping and viral load) in clinical samples. Methods Published HCV sequences were compared to select specific primers, probe and restriction enzyme sites. An original real-time nested RT-PCR-RFLP assay was then developed and validated to detect, genotype and quantify HCV in plasma samples. Results The real-time nested RT-PCR data were linear and reproducible for HCV analysis in clinical samples. High correlations (> 0.97) were observed between samples with different viral loads and the corresponding read cycle (Ct - Cycle threshold), and this part of the assay had a wide dynamic range of analysis. Additionally, HCV genotypes 1, 2 and 3 were successfully distinguished using the RFLP method. Conclusions A complete serial molecular assay was developed and validated for HCV detection, quantification and genotyping. .


Subject(s)
Humans , /genetics , Hepacivirus/genetics , Hepatitis C/diagnosis , RNA, Viral/blood , DNA Primers , Genotype , Hepacivirus/isolation & purification , Polymorphism, Restriction Fragment Length , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity , Viral Load
14.
Mem. Inst. Oswaldo Cruz ; 108(3): 392-394, maio 2013.
Article in English | LILACS | ID: lil-676978

ABSTRACT

The neuraminidase (NA) genes of A(H1N1)pdm09 influenza virus isolates from 306 infected patients were analysed. The circulation of oseltamivir-resistant viruses in Brazil has not been reported previously. Clinical samples were collected in the state of Rio Grande do Sul (RS) from 2009-2011 and two NA inhibitor-resistant mutants were identified, one in 2009 (H275Y) and the other in 2011 (S247N). This study revealed a low prevalence of resistant viruses (0.8%) with no spread of the resistant mutants throughout RS.


Subject(s)
Humans , Antiviral Agents/pharmacology , Drug Resistance, Viral/genetics , Influenza A Virus, H1N1 Subtype/drug effects , Mutation , Neuraminidase/genetics , Oseltamivir/pharmacology , Brazil , Influenza A Virus, H1N1 Subtype/enzymology , Influenza A Virus, H1N1 Subtype/genetics , Microbial Sensitivity Tests , Reverse Transcriptase Polymerase Chain Reaction , RNA, Viral/genetics
15.
Braz. j. microbiol ; 44(2): 505-510, 2013. tab
Article in English | LILACS | ID: lil-688586

ABSTRACT

Mycoplasma gallisepticum (MS) and Mycoplasma synoviae (MS) are important avian pathogens and cause economic losses to the poultry industry. Molecular biology techniques are currently used for a rapid detection of these pathogens and the adoption of control measures of the diseases. The aim of this study was to develop and validate a technique for simultaneous detection of MG and MS by multiplex real time polymerase chain reaction (PCR). The complete assay (Multiplex MGMS) was designed with primers and probes specific for each pathogen and developed to be carried out in a single tube reaction. Vaccines, MG and MS isolates and DNA from other Mycoplasma species were used for the development and validation of the method. Further, 78 pooled clinical samples from different poultry flocks in Brazil were obtained and used to determine the sensitivity and specificity of the technique in comparison to 2 real time PCR assays specific for MG (MG PCR) and MS (MS PCR). The results demonstrated an agreement of 100% (23 positive and 44 negative samples) between Multiplex MGMS and MG PCR in the analysis of 67 samples from MG positive and negative poultry flocks, and an agreement of 96.9% between Multiplex MGMS and MS PCR in the analysis of 64 samples from MS positive and negative poultry flocks. Considering the single amplification tests as the gold standard, the Multiplex MGMS showed 100% of specificity and sensitivity in the MG analysis and 94.7% sensitivity and 100% specificity in the MS analysis. This new assay could be used for rapid analysis of MG and MS in the poultry industry laboratories.


Subject(s)
Animals , Molecular Diagnostic Techniques/methods , Mycoplasma Infections/veterinary , Mycoplasma gallisepticum/isolation & purification , Mycoplasma synoviae/isolation & purification , Poultry Diseases/diagnosis , Real-Time Polymerase Chain Reaction/methods , Veterinary Medicine/methods , Brazil , Bacteriological Techniques/methods , Multiplex Polymerase Chain Reaction/methods , Mycoplasma Infections/diagnosis , Mycoplasma Infections/microbiology , Mycoplasma gallisepticum/genetics , Mycoplasma synoviae/genetics , Poultry , Poultry Diseases/microbiology , Sensitivity and Specificity
16.
Braz. j. infect. dis ; 15(5): 467-472, Sept.-Oct. 2011. ilus, tab
Article in English | LILACS | ID: lil-612706

ABSTRACT

BACKGROUND: It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. OBJECTIVES: Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. METHODS: Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. RESULTS: The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9 percent) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5 percent increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. CONCLUSION: The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.


Subject(s)
Female , Humans , Cervix Uteri/virology , Papillomaviridae/genetics , Papillomavirus Infections/diagnosis , Base Sequence , DNA Primers/analysis , DNA, Viral/analysis , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Papillomaviridae/classification , Papillomavirus Infections/virology , Sensitivity and Specificity
17.
Rev. saúde pública ; 44(6): 1094-1101, dez. 2010. ilus, tab
Article in Portuguese | LILACS | ID: lil-565096

ABSTRACT

OBJETIVO: Estimar a prevalência dos subtipos do HIV-1 e analisar fatores associados. MÉTODOS: Foi realizado um estudo transversal com amostra de conveniência de 80 pacientes adultos HIV-positivos atendidos em serviço especializado em DST/Aids em Canoas, RS, no período de julho de 2008 a janeiro de 2009. A determinação dos subtipos do HIV foi realizada por amplificação de fragmento do genoma viral pela reação em cadeia da polimerase seguida do seqüenciamento dos fragmentos amplificados. Variáveis sociodemográficas, clínicas e comportamentais foram coletadas em questionário estruturado. Foi realizada análise estatística univariada utilizando os testes de qui-quadrado e t de Student. RESULTADOS: Foi observada uma prevalência maior do subtipo C (43,8 por cento; IC 95 por cento: 32,9;54,6), seguida pelo CRF31_BC (35,0 por cento; IC 95 por cento: 24,6;45,5) e subtipos B (18,8 por cento; IC 95 por cento: 10,2;27,3) e F (2,4 por cento; IC 95 por cento: 0;5,9). Outros subtipos de HIV-1 não foram observados. Pacientes infectados com CRF31_BC apresentaram diagnóstico mais recente do que os pacientes infectados com o subtipo B (p < 0,05). Observou-se também maior freqüência de co-infecção com outros vírus (hepatites B e C e T-linfotrópicos humanos) nos indivíduos portadores do CRF31_BC do que nos demais subtipos. Com relação aos aspectos sociodemográficos, não foram observadas diferenças na distribuição dos subtipos e formas recombinantes quanto ao sexo e práticas sexuais. CONCLUSÕES: Os resultados obtidos indicam uma freqüência maior do subtipo C e do CRF31_BC nesse centro urbano do sul do Brasil, com possíveis vias de transmissão diferentes.


OBJECTIVE: To estimate the prevalence of HIV-1 subtypes and analyze factors associated. METHODS: A cross-sectional study was performed with a convenience sample of 80 adult HIV-positive patients, users of an AIDS/STD specialized service, in the city of Canoas, Southern Brazil, between July 2008 and January 2009. Determination of HIV subtypes was performed with the amplification of viral genome fragment, using polymerase chain reaction, followed by sequencing of the amplified fragments. Sociodemographic, clinical and behavioral variables were collected in a structured questionnaire. Univariate statistical analysis was performed, using chi-square test and Student's t-test. RESULTS: A higher prevalence of subtype C was found (43.8%; 95% CI: 32.9;54.6), followed by CRF31_BC (35.0%; 95% CI: 24.6;45.5) and subtypes B (18.8%; 95% CI: 10.2;27.3) and F (2.4%; 95% CI: 0;5.9). Other HIV-1 subtypes were not observed. Patients infected with CRF31_BC were diagnosed more recently than patients infected with subtype B (p<0.05). In addition, there was a higher frequency of co-infection with other viruses (hepatitis B and C and human T-lymphotropic viruses) in individuals with CRF31_BC, compared to other subtypes. With regard to sociodemographic aspects, there were no differences in the distribution of subtypes and recombinant forms, in terms of gender and sexual practices. CONCLUSIONS: Results obtained indicate a higher frequency of subtype C and CRF31_BC in this urban center of Southern Brazil, with possible different ways of transmission.


OBJETIVO: Estimar la prevalencia de los subtipos de VIH-1 y analizar factores asociados. MÉTODOS: Se realizó un estudio transversal con muestra de conveniencia de 80 pacientes adultos VIH-positivos atendidos en servicio especializado en DST/Sida en Canoas, Sur de Brasil, en el período de julio de 2008 a enero de 2009. La determinación de los subtipos de VIH fue realizada por amplificación de fragmento del genoma viral por la reacción en cadena de la polimerasa seguida de la secuenciación de los fragmentos amplificados. Variables sociodemográficas, clínicas y conductuales fueron colectadas en cuestionario estructurado. Se realizó análisis estadístico univariado utilizando las pruebas de chi-cuadrado y t de Student. RESULTADOS: Se observó una prevalencia mayor del subtipo C (43,8%; IC 95%:32,9;54,6), seguido de la CRF31_BC (35,0%; IC 95%: 24,6;45,5) y subtipos B (18,8%; IC 95%: 10,2;27,3) y F (2,4%; IC 95%: 0;5,9). Otros subtipos de VIH-1 no fueron observados. Pacientes infectados con CRF31_BC presentaron diagnóstico más reciente que los pacientes infectados con el subtipo B (p<0,05). Se observó también una mayor frecuencia de co-infección con otros virus (hepatitis B y C y T-linfotrópicos humanos) en los individuos portadores del CRF31_BC en comparación con los demás subtipos. Con relación a los aspectos sociodemográficos, no fueron observadas diferencias en la distribución de los subtipos y formas recombinantes con relación al sexo y prácticas sexuales. CONCLUSIONES: Los datos obtenidos en el presente estudio indican una mayor frecuencia del subtipo C y de la CRF31_BC en este centro urbano del sur de Brasil y sugieren que dichos subtipos pueden presentar vías de transmisión diferentes.


Subject(s)
HIV Seroprevalence , HIV Infections/epidemiology , Acquired Immunodeficiency Syndrome/epidemiology , Cross-Sectional Studies
18.
Article in Portuguese | LILACS | ID: lil-566981

ABSTRACT

A hepatite B pode ser classificada em oito diferentes genótipos (A-H). Esses genótipos diferem na sua distribuição geográfica mundial. No Brasil, os genótipos mais freqüentemente encontrados são o A, D e F. Algumas alterações na estrutura genética desses genótipos podem resultar em diferentes níveis de patogenicidade, sendo relacionadas com maior ou menor risco de desenvolvimento de hepatocarcinoma ou cirrose no fígado. Além das diferenças citadas, a heterogeneidade dos genótipos da hepatite B parece estar relacionada com diferenças na evolução clínica da infecção e na resposta ao tratamento antiviral. Alguns genótipos demonstraram responder melhor ao tratamento com interferon ou nucleotídeos análogos do que outros. O objetivo desta revisão foi demonstrar a importância do tratamento da hepatite B baseado nos seus diferentes genótipos. Foram revisados artigos da literatura, selecionando aqueles que abordavam questões relacionadas aos genótipos da hepatite B e sua relação com o tratamento desta infecção. Nos artigos revisados, o tratamento da hepatite B baseada em genótipos apresentou diferenças significativas. Os genótipos A e B parecem ter uma melhor resposta ao tratamento antiviral com interferon alfa e/ou lamivudina; porém, mais estudos são necessários para a consistência dessa afirmação. No entanto, através dos presentes dados, já é possível demonstrar forte associação entre genótipos e resposta antiviral. Deste modo, adaptar o tratamento aos genótipos pode promover uma melhor resposta do interferon e dos nucleotídeos na terapêutica da infecção pelo vírus da hepatite B.


Type B hepatitis can be classified according to eight different genotypes (A-H). These genotypes are different in terms of worldwide geographical distribution. In Brazil the most frequent genotypes are A, D and F. Some changes in the genetic structure of these genotypes can cause different levels of pathogenesis, being related to lower or higher risk of developing hepatocellular carcinoma or liver cirrhosis. In addition to the above mentioned differences, heterogeneity of hepatitis B genotypes seems to be related to the differences in clinical evolution of the infection and response to antiviral treatment. Some genotypes proved to have a better response to the treatment using interferon or similar nucleotides than others. This review aimed at showing the importance of treatment of type B hepatitis based on its different genotypes. Different articles from the specific medical literature were reviewed and those including genotypes for type B hepatitis and their association with the treatment of this infection were selected. In the reviewed articles genotypebased treatment of hepatitis B showed significant differences. Genotypes A and B seem to have a better response to the antiviral treatment with alpha interferon and/or lamivudine; however, more studies are necessary to confirm this assertion. Nevertheless, using the present data it is already possible to prove a strong connection between genotypes and antiviral response. Therefore, adjusting treatment to genotypes can cause a better therapeutic response from interferon and nucleotides in type B hepatitis therapy.


Subject(s)
Humans , Hepatitis B/etiology , Hepatitis B/genetics , Hepatitis B/therapy , Genotype , Interferon-alpha/therapeutic use , Lamivudine/therapeutic use
19.
Acta bioquím. clín. latinoam ; 33(3): 371-4, sept. 1999. tab
Article in Portuguese | LILACS | ID: lil-258463

ABSTRACT

A reaçao em cadeira da polimerase (PCR-Polymerase Chain Reaction) é uma técnica de amplificaçao enzimática de seqüências específicas de ácidos nucleicos. Essa técnica tem sido amplamente descrita para detecçao e tipagem do Papilomavírus Humano (HPV - Human Papilomavirus). Neste trabalho, 460 amostras de colo uterino foran avaliadas para a presença do DNA do HPV pela técnica de PCR. Amostras positivas foram subseqüentemente tipadas por RFLP (Restriction Fragment Lenght Polymorphism). Os resultados de PCR-RFLP foram comparados com os exames colposcópico e citopatológico (Papanicolou). O PCR-RFLP demonstrou ser uma técnica eficaz na detecçao e tipagem virais, apresentando maior sensibilidade do que os exames citopatológico e colposcópico


Subject(s)
Humans , Female , Papillomaviridae/isolation & purification , Papillomavirus Infections/diagnosis , Cervix Uteri/virology , Colposcopy , Polymerase Chain Reaction , Risk Factors , Vaginal Smears
20.
Rev. bras. anal. clin ; 29(4): 203-204, 1997. tab
Article in Portuguese | LILACS | ID: lil-525248

ABSTRACT

A Reação em Cadeia da Polimerase (PCR – Polymerase Chain Reaction) é uma técnica de amplificação enzimática de sequências específicas de ácidos nucléicos. Essa técnica tem sido amplamente descrita para detecção e tipagem do Papilomavírus Humano (HPV – Human Papillomavirus). Neste trabalho, 460 amostras de colo uterino foram avaliadas para a presença do DNA do HPV pela técnica de PCR. Amostras positivas foram subsequentemente tipadas por RFLP (Restriction Fragment Polymorphism). Os resultados de PCR-RFLP foram comparados com os exames colposcópico e citopatológico (Papanicolaou). O PCR-RFLP demonstrou ser uma técnica eficaz na detecção e tipagem virais, apresentando maior sensibilidade do que os exames citopatológico e colposcópico.


Subject(s)
Humans , Female , Bacterial Typing Techniques , Clinical Laboratory Techniques , DNA Probes, HPV , Papillomavirus Infections , Polymerase Chain Reaction
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