COMPARAÇÃO DE MÉTODOS PARA EXTRAÇÃO DE DNA NA REAÇÃO EM CADEIA DA POLIMERASE PARA DETECÇÃO DE Salmonella enterica SOROVAR Enteritidis EM PRODUTOS AVÍCOLAS

Filho, Raphael Lúcio Andreatti; Marietto-Gonçalves, Guilherme Augusto; Okamoto, Adriano Sakai; Lima, Edna Tereza de

doi:10.5216/cab.v12i1.3774

Cited by 9 publications

(2 citation statements)

References 3 publications

(3 reference statements)

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“…According to Monteiro, Montanhini andBittencourt (2014) &Shetty, Ghosh andPaul (2017) between the two extraction methods used in their research, CTAB and thermal lysis, it was proven that the best method of extraction was by thermal lysis, this method proved to be effective for certain Gram-positive bacteria, which has a thick layer of glycan peptide resulting in better extraction in terms of quantity and purity. However, Andreatti Filho et al (2011) also used the same methods to extract the DNA of a certain Gramnegative bacterium in their research and analyzed that between two tests, both extractions had satisfactory quantity, quality and purity of genetic material, CTAB was the one that showed high sensitivity and quality with good clarity in the bands visualized.…”

Section: Discussionmentioning

confidence: 99%

Optimization of Gram Positive Bacteria Dna Extraction Protocols and Purity and Concentration Analysis by Gel Electrophoresis

Alves¹,

Silva

Lopes³

et al. 2021

Rev. Interfaces

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This work aims to optimize protocols for extraction of bacterial Deoxyribonucleic Acid (DNA) in order to develop more practical, fast and less costly methodologies, therefore bringing improvements to the development of research and applications in laboratory routine based on the difficulty of establishing an ideal method that can extract Grampositive bacterial DNA with excellent quality. The material and the bacterial strain Staphylococcus aureus 15 were provided by the Leão Sampaio University Center, where tests for DNA extraction were also carried out. The tests used were Sodium Dodecyl Sulfate (SDS); salting-out; PROMEGA's Kit and CTAB / Phenol / Chloroform and were analyzed using agarose gel electrophoresis. The best results were obtained through the use of PROMEGA's commercial kit, salting-out without the use of Proteinase K and CTAB / Phenol / Chloroform, with salting-out having greater advantages in relation to other methods because it brings speed, low cost, good concentration of extracted material and not be toxic. The molecular tests for the extraction of Staphyloccocus aureus DNA were successful in general, when comparing issues such as low cost, extracted concentration, sharpness of the bands in electrophoresis and practicality of the performance. The best method for a small laboratory routine or for future research would be the salting-out without proteinase K, thus concluding that it is possible to obtain an excellent DNA extraction with less complexity and low cost methods.

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Section: Discussionmentioning

confidence: 99%

Optimization of Gram Positive Bacteria Dna Extraction Protocols and Purity and Concentration Analysis by Gel Electrophoresis

Alves¹,

Silva

Lopes³

et al. 2021

Rev. Interfaces

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show abstract

“…All samples identified as E. coli were subjected to DNA extraction by thermal method (Andreatti Filho et al, 2011) for the detection of eae, stx 1 , stx 2, and bfp virulence genes for PCR using pairs of specific primers for each gene (Table 1).…”

Section: Pcrmentioning

confidence: 99%

Detection and Antimicrobial Resistance Profile of Enteropathogenic (EPEC) and Shigatoxigenic Escherichia coli (STEC) in Conventional and Organic Broiler Chickens

Lopes,

Alves,

Costa

et al. 2023

Braz. J. Poult. Sci.

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Enteropatogenic Escherichia coli (EPEC) and shigatoxigenic E. coli (STEC), are generally poultry and poultry product isolate and can cause serious human infections. Many strains may become resistant to various antimicrobials, which can hinder the treatment of bacterial diseases. Organic farming seeks to avoid the selection and frequency of antimicrobial-resistant bacteria. This study aims to verify the resistance of EPEC and STEC from organic and conventional (industrial) broiler isolates to antimicrobials. All isolates were submitted to disk diffusion test with tetracycline, gentamicin, enrofloxacin, ceftriaxone and amoxicillin + clavulanate (TET, GEN, ENO, CTX, AMC) and PCR to detect specific virulence genes for EPEC and STEC. A total of 297 E. coli strains were isolated, 213 from conventional. In organic broiler, 84 strains were isolated. The strains from the conventional broiler isolates were resistant to five antimicrobials tested: TET 48.82% (104/213), ENO 28.17% (60/213), CTX 15.49% (33/213), GEN 14.55% (31/213), and AMC 7.04% (15/213), and 9.86% (21/213) were considered multidrug-resistant. Organic chicken strains were resistant to four of the antimicrobials tested: TET 35.7% (30/84), ENO 9.5% (8/84), CTX 2.4% (2/84), GEN 4.8% (4/84). Of the strains from the organic broiler chicken isolates, only 1.2% (1/84) was considered multidrug-resistant. No EPEC and STEC were found in the organic chicken samples. The multidrug resistance was characterized in 9.52% (2/21) of the EPEC and 4.76% (1/21) of the STEC. The study demonstrated the absence of EPEC and STEC strains in organic broilers and carcasses and a lower frequency of multiresistant strains compared to conventional breeding.

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Comparison of three diagnostic methods for Salmonella enterica serovars detection in chicken rinse

et al. 2018

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Salmonella detection is a key point in food safety testing, because of the frequent association of this pathogen with food poisoning in humans. The standard bacteriological tests currently used for Salmonella-detection are time-consuming; therefore, there is a need to develop alternative methods to accelerate the detection. In order to accelerate Salmonella diagnosis, we used the immunomagnetic separation assay associated with bacteriophage P22 for the rapid detection of the following Salmonella serovars in chicken rinses of drumsticks, artificially contaminated with 5, 10, and 100 CFU/25mL of bacteria: Salmonella enterica subsp. enterica serovar Heidelberg (S. Heidelberg), Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium). The efficiency of the technique, represented by the time required for detection of positive and negative samples, was compared with that of the standard diagnostic tests used for this pathogen, the bacteriological assay and the polymerase chain reaction (PCR)-based test. This study confirmed the ability of the bacteriophage-associated immunomagnetic separation assay to identify 99.6% of Salmonella-positive samples of the three serovars tested. In contrast, the bacteriological assay and PCR-based test detected 95.1% and 98.5% of the Salmonella-positive samples respectively.

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COMPARAÇÃO DE MÉTODOS PARA EXTRAÇÃO DE DNA NA REAÇÃO EM CADEIA DA POLIMERASE PARA DETECÇÃO DE Salmonella enterica SOROVAR Enteritidis EM PRODUTOS AVÍCOLAS

Cited by 9 publications

References 3 publications

Optimization of Gram Positive Bacteria Dna Extraction Protocols and Purity and Concentration Analysis by Gel Electrophoresis

Optimization of Gram Positive Bacteria Dna Extraction Protocols and Purity and Concentration Analysis by Gel Electrophoresis

Detection and Antimicrobial Resistance Profile of Enteropathogenic (EPEC) and Shigatoxigenic Escherichia coli (STEC) in Conventional and Organic Broiler Chickens

Comparison of three diagnostic methods for Salmonella enterica serovars detection in chicken rinse

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