SABIIA - Sistema Aberto e Integrado de Informação em Agricultura (Sabiia)

Caballero,O.L.; Costa,M.C.S.L.; Trevisan,A.; Oliveira,R.M.; Viotti,E.A.; Távora,E.R.F.; Vilaça,S.S.; Sabagga,E.; de-Paula,F.J.; Távora,P.F.; Brasileiro-Filho,G.; Villa,L.L.; Simpson,A.J.G..

To assess the clinical relevance of a semi-quantitative measurement of human cytomegalovirus (HCMV) DNA in renal transplant recipients within the typical clinical context of a developing country where virtually 100% of both receptors and donors are seropositive for this virus, we have undertaken HCMV DNA quantification using a simple, semi-quantitative, limiting dilution polymerase chain reaction (PCR). We evaluated this assay prospectively in 52 renal transplant patients from whom a total of 495 serial blood samples were collected. The samples scored HCMV positive by qualitative PCR had the levels of HCMV DNA determined by end-point dilution-PCR. All patients were HCMV DNA positive during the monitoring period and a diagnosis of symptomatic infection was...

Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method

Palavras-chave: Polymerase chain reaction; Human cytomegalovirus; Renal transplant; Diagnosis; Viral load.

Ano: 1999

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999001200010

Li,Chao; Fu,Gongyu; Shi,Yaoqiang; Zhang,A-Mei; Xia,Xueshan; Fang,Yue; Mao,Xiaoqin; Jiang,Jie; Song,Yuzhu; Yang,Guangying.

Klebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of K. pneumoniae-specific gene ureR_1 (Gene ID: 11847803). The ureR_1 gene was obtained through local and online BLAST, and the specific primers were designed for its detection. Positive reactions were observed on all 140 K. pneumoniae clinical isolates while all the 82 non-K. pneumoniae clinical isolates were negative. Plasmids with the specific gene and the mouse...

DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells

Palavras-chave: Loop-mediated isothermal amplification; Polymerase chain reaction; Klebsiella pneumoniae; Novel specific gene; Specific; Sensitive method.

Ano: 2019

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2019000300609

Ribeiro,C.N.M.; Peres,L.C.; Pina-Neto,J.M..

The objective of the present study was to develop a simplified low cost method for the collection and fixation of pediatric autopsy cells and to determine the quantitative and qualitative adequacy of extracted DNA. Touch and scrape preparations of pediatric liver cells were obtained from 15 cadavers at autopsy and fixed in 95% ethanol or 3:1 methanol:acetic acid. Material prepared by each fixation procedure was submitted to DNA extraction with the Wizard® genomic DNA purification kit for DNA quantification and five of the preparations were amplified by multiplex PCR (azoospermia factor genes). The amount of DNA extracted varied from 20 to 8,640 µg, with significant differences between fixation methods. Scrape preparation fixed in 95% ethanol provided...

A polymerase chain reaction-based assay to identify genotype F of hepatitis B virus

Palavras-chave: Autopsy; Congenital anomalies; DNA; Touch preparative; Scrape preparative; Polymerase chain reaction.

Ano: 2004

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000500002

Comparison of the diagnosis of malaria by microscopy, immunochromatography and PCR in endemic areas of Venezuela

Moraes,M.T.B.; Niel,C.; Gomes,S.A..

We have developed a polymerase chain reaction (PCR) assay which distinguishes genotype F from the other genotypes of hepatitis B virus (HBV). The method was used to characterize HBV strains isolated in urban areas of the Brazilian Amazon. DNA was amplified in 54 of a total of 78 HBsAg-positive serum samples, using universal, non-genotype-specific primers. Only 4 (7.4%) were identified as genotype F by our genotype-specific PCR assay. This proportion is notably lower than that previously reported in Argentina, Venezuela, Peru, and Central America.

Tipo: Info:eu-repo/semantics/other

Palavras-chave: Hepatitis B virus; Genotype F; Subtype adw4; Polymerase chain reaction; Brazil.

Ano: 1999

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000100006

Rodulfo,H; De Donato,M; Mora,R; González,L; Contreras,C.E.

Whole blood samples (N = 295) were obtained from different locations in Amazonas and Sucre States, in Venezuela. Malaria was diagnosed by microscopy, OptiMAL™ and polymerase chain reaction (PCR), with Plasmodium vivax, P. falciparum, and P. malariae being detected when possible. We identified 93 infections, 66 of which were caused by P. vivax, 26 by P. falciparum, and 1 was a mixed infection. No infection caused by P. malariae was detected. The sensitivity and specificity of each diagnostic method were high: 95.7 and 97.9% for microscopy, 87.0 and 97.9% for OptiMAL, and 98.0 and 100% for PCR, respectively. Most samples (72.2%) showed more than 5000 parasites/µL blood. The sensitivity of the diagnosis by microscopy and OptiMAL decreased with lower...

PCR identification of Mycobacterium tuberculosis complex in a clinical sample from a patient with symptoms of tuberculous spondylodiscitis

Palavras-chave: Malaria; Molecular diagnosis of malaria; Microscopic diagnosis of malaria; Polymerase chain reaction; OptiMAL.

Ano: 2007

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2007000400012

Absence of hepatitis B virus DNA in patients with hepatitis C and non-A-E hepatitis in the State of São Paulo, Brazil

Ribeiro,M.A.; Barouni,A.S.; Augusto,C.J.; Augusto,M.V.S.; Lopes,M.T.P.; Salas,C.E..

A 42-year-old male complaining of thoracic spine pain was admitted to the hospital for evaluation. An X-ray and computer tomography of the thoracic spine showed spondylodiscitis of the L3 lumbar and L2-L3 intervertebral disk. The tuberculin skin test (PPD) was strongly positive. A radioscopy-guided fine needle aspirate of the affected area was cultured but did not reveal the cause of the disease. Two biopsy attempts failed to reveal the cause of the disease by culturing or by acid-fast-resistant staining (Ziehl Neelsen) of the specimens. A third biopsy also failed to detect the infectious agent by using microbiological procedures, but revealed the presence of a 245-bp amplicon characteristic of the Mycobacterium tuberculosis complex after PCR of the...

Tipo: Info:eu-repo/semantics/report

Palavras-chave: Spondylodiscitis; Tuberculosis; Polymerase chain reaction; Mycobacterium tuberculosis complex.

Ano: 2007

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2007000100001

Risk of hepatitis B virus transmission by diagnostic hysteroscopy

Souza,L.O.; Pinho,J.R.R.; Carrilho,F.J.; da Silva,L.C..

Occult hepatitis B virus (HBV) infection has been reported as cases in which HBV DNA was detected despite the absence of any HBV serological markers or in cases in which anti-HBc antibody was the sole marker. The aim of the present study was to determine, using the polymerase chain reaction (PCR), whether HBV infection occurs in hepatitis C and non-A-E hepatitis patients without serological evidence of hepatitis B infection in São Paulo State. Two different populations were analyzed: 1) non-A-E hepatitis patients, including 12 patients with acute and 50 patients with chronic hepatic disorders without serological evidence of infection with known hepatitis viruses; 2) 43 patients previously diagnosed as hepatitis C with positive results for anti-HCV and HCV...

Tipo: Info:eu-repo/semantics/other

Palavras-chave: Hepatitis B virus; Non-A-E hepatitis; Polymerase chain reaction.

Ano: 2004

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004001100011

Santos,N.C.; Pinho,J.R.R.; Lemos,M.F.; Moreira,R.C.; Lopes,C.M.C.; Sacilotto,M.T.J.; Tacla,M.; Pinheiro,W.S.; Ramos,L.O..

Few data are available in the literature concerning the efficacy of standard hysteroscope disinfection procedures to prevent hepatitis B transmission. The aim of the present study was to determine the risk of hepatitis B virus (HBV) transmission during hysteroscopy among anti-HBc-seropositive women. Serum and hysteroscopic samples were collected from 62 women after diagnostic hysteroscopy. All samples were tested for serologic HBV markers. Polymerase chain reactions (PCR) were carried out to amplify regions C and S of the viral genome and only samples amplified by both pairs of primers were considered to be positive. Anti-HBc was repeatedly reactive in 48 (77%) of 62 serum samples, and HBsAg was detected in 8 (13%). At least one HBV serologic marker was...

Kaposi's sarcoma-associated herpesvirus infection and Kaposi's sarcoma in Brazil

Palavras-chave: Hepatitis B; Hysteroscopy; Disinfection; Polymerase chain reaction.

Ano: 2004

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000500009

Ramos-da-Silva,S.; Elgui-de-Oliveira,D.; Borges,L.; Bacchi,C.E..

Kaposi's sarcoma (KS) became a critical health issue with the emergence of acquired immunodeficiency syndrome (AIDS) in the 1980s. Four clinical-epidemiological forms of KS have been described: classical KS, endemic KS, iatrogenic KS, and AIDS-associated KS. In 1994, Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus type 8 was identified by Chang and colleagues, and has been detected worldwide at frequencies ranging from 80 to 100%. The aim of the present study was to evaluate the frequency of KSHV infection in KS lesions from HIV-positive and HIV-negative patients in Brazil, as well as to review the current knowledge about KS transmission and detection. For these purposes, DNA from 51 cases of KS was assessed by PCR: 20 (39.2%) cases of...

Acute and latent infection in mice with a virulent strain of Aujeszkys disease virus

Palavras-chave: Kaposi's sarcoma; Human herpesvirus type 8; Kaposi's sarcoma-associated herpesvirus; Polymerase chain reaction; Viral carcinogenesis; Brazil.

Ano: 2006

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000500002

Flatschart,Roberto B.; Resende,Maurício.

Acute and latent infections with the Brazilian LA031 strain of Aujeszkys disease virus (ADV) were established in mice. Ultraviolet irradiated ADV administered subcutaneously was a successful way to establish latent infection. The presence of ADV was detected by PCR. Two sets of 22-mer primers were synthesized and used to amplify gG glycoprotein gene sequences in acute and latent infected trigeminal nerve ganglia. The specificity of the amplification was verified by dot-blot hybridization.

Prevalence of virulence genes among bulgarian nosocomial and cystic fibrosis isolates of Pseudomonas aeruginosa

Palavras-chave: Aujeszkys disease virus; Polymerase chain reaction; Dot-blot hybridization.

Ano: 2000

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822000000400013

Mitov,Ivan; Strateva,Tanya; Markova,Boyka.

The aim of this study was to evaluate the prevalence of some virulence genes among 202 Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients (n=42) and non-CF in-patients (n=160) and to analyze the values according to the patient groups, infection localization and antimicrobial resistance. The following frequencies in all studied strains were established: algD (encoding GDP-mannose 6-dehydrogenase AlgD) - 91.1%, pilB (type IV fimbrial biogenesis protein PilB) - 23.8%, nan1 (neuraminidase) - 21.3%, lasB (elastase LasB) - 100%, plcH (haemolytic phospholipase C precursor) - 91.6%, exoS (exoenzyme S) - 62.4%, and exoU (exoenzyme U) - 30.2%. The prevalence of nan1 was significantly higher (P<0.01) in CF isolates (38.1%) than that in non-CF...

Investigation of the association between clinical outcome and the cag pathogenicity-island and other virulence genes of Helicobacter pylori isolates from patients with dyspepsia in Eastern Turkey

Palavras-chave: Pseudomonas aeruginosa; Virulence genes; Polymerase chain reaction; Cystic fibrosis isolates; Non-cystic fibrosis isolates.

Ano: 2010

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000300008

Ozbey,Gokben; Demirel,Ulvi; Aygun,Cem; Ertas,Hasan Basri.

The aims of our work were to determine the presence of the cag pathogenicity-island (cag PAI) and other virulence genes of Helicobacter pylori recovered from patients with gastritis and peptic ulcer, and to investigate the correlation of these virulence genes with clinical outcome. The presence of the cagA, the promoter regions of cagA, cagE, cagT, and the left end of cag-PAI (LEC), cag right junction (cagRJ), the plasticity region open reading frames (ORFs), vacA and oipA genes among 69 H. pylori isolates were determined by polymerase chain reaction. Intact cag PAI was detected in only one (1.4%) isolate. The cagA gene was identified in 52.1% and 76.2% of isolates from patients with dyspepsia (gastritis and peptic ulcer), respectively. The plasticity...

Development of a propidium monoazide-polymerase chain reaction assay for detection of viable Lactobacillus brevis in beer

Palavras-chave: Helicobacter pylori; Gastritis; Peptic ulcer; Cag pathogenicity-island; Polymerase chain reaction.

Ano: 2013

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000400034

Ma,Yanlin; Deng,Yang; Xu,Zhenbo; Liu,Junyan; Dong,Jianjun; Yin,Hua; Yu,Junhong; Chang,Zongming; Wang,Dongfeng.

ABSTRACT The spoilage of beer by bacteria is of great concern to the brewer as this can lead to turbidity and abnormal flavors. The polymerase chain reaction (PCR) method for detection of beer-spoilage bacteria is highly specific and provides results much faster than traditional microbiology techniques. However, one of the drawbacks is the inability to differentiate between live and dead cells. In this paper, the combination of propidium monoazide (PMA) pretreatment and conventional PCR had been described. The established PMA-PCR identified beer spoilage Lactobacillus brevis based not on their identity, but on the presence of horA gene which we show to be highly correlated with the ability of beer spoilage LAB to grow in beer. The results suggested that...

The application of PCR in the detection of mycotoxigenic fungi in foods

Palavras-chave: Propidium monoazide; Polymerase chain reaction; Beer spoilage bacteria; Lactobacillus brevis horA.

Ano: 2017

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000400740

Konietzny,Ursula; Greiner,Ralf.

It is estimated that 25 to 50% of the crops harvested worldwide are contaminated with mycotoxins. Because of the toxic and carcinogenic potential of mycotoxins, there is an urgent need to develop detection methods that are rapid and highly specific. The highly advanced physico-chemical methods for the analysis of mycotoxins in use, have the disadvantage that highly sophisticated clean-up and/or derivatization procedures must be applied. An alternative could be the detection of the mycotoxigenic moulds themselves, especially as molecular techniques have been introduced recently as powerful tools for detecting and identifying fungi. PCR methods for the detection of aflatoxigenic Aspergilli, patulin-producing Penicillum and trichothecene- as well as...

Clinical and Laboratory evaluation of measleslike rash in children and young adults

Palavras-chave: Aflatoxin; Fumonisin; Patulin; Polymerase chain reaction; Trichothecene.

Ano: 2003

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000400001

Stewien,Klaus Eberhard; Lima,Lourdes Rehder de Andrade Vaz de; Botosso,Viviane Fongaro; Oliveira,Maria Isabel de; Fagundes,Simone N.; Nogueira,Meri B.; Ragazzi,Selma Lopes Betta; Costa,Maria Tereza Zuluni da; Ejzenberg,Bernardo; Durigon,Edison Luiz.

A clinical and laboratory evaluation of 11 children and young adults with measleslike rash was done during the measles outbreak in the Greater São Paulo Metropolitan area at the end of 1996 and spread over the country during 1997. Measles was laboratory confirmed in 07 patients by specific IgM detection in acute serum specimens using an IgM-capture EIA, by specific IgG seroconversion in serum pairs, and by reverse transcription PCR and virus isolation in peripheral blood lymphocytes. Clinical presentations were not always classic; one of the 07 cases had received measles vaccine and corresponded to modified clinical case of measles. The 4 remaining cases were negative for measles and were diagnosed as exanthem subitum (2 cases), scarlet fever and Kawasaki...

A quantitative analysis of Propionibacterium acnes in lesional and non-lesional skin of patients with progressive macular hypomelanosis by real-time polymerase chain reaction

Palavras-chave: Measles; Clinical evaluation; Laboratory investigation; Polymerase chain reaction; Measles virus isolation; Enzyme immunoassay.

Ano: 2000

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822000000400008

Cavalcanti,Silvana Maria de Morais; França,Emmanuel Rodrigues de; Magalhães,Marcelo; Lins,Ana Kelly; Brandão,Laura Costa; Magalhães,Vera.

Little is known about the etiology of progressive macular hypomelanosis, although it has been suggested that Propionibacterium acnes plays an important role. While microbiological culture is commonly employed to identify Propionibacterium acnes, new identification methods have been under investigation, amongst them polymerase chain reaction. To determine the cut-off point for the number of genome copies of Propionibacterium acnes in the lesional skin of patients with progressive macular hypomelanosis as a positive marker, employing quantitative real-time polymerase chain reaction and anaerobic culture, considered gold standard. An observational study with a comparison group, included 35 patients with dermatosis, attended at the Oswaldo Cruz University...

Prevalence of hemoglobinopathies in school children: the importance of using confirmatory methods

Palavras-chave: Hypopigmentation/progressive macular hypomelanosis; Propionibacterium acnes; Polymerase chain reaction; Biopsy/bacteriology; ROC curve.

Ano: 2011

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000200002

BJPS

Tavares,Cristiane Fernandes de Freitas; Guimarães,Jacqueline da Silva; Souza,Ana Maria de.