Comparative Study of Rapid DNA Extraction Methods of Pathogenic Bacteria

Abdelhai, Mandour H.

doi:10.11648/j.bio.20160401.11

Cited by 23 publications

(21 citation statements)

References 23 publications

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“…DNA was extracted by the boiling method (Mandour et al ) from one cefotaxime‐resistant E. coli of each positive sample. Standard PCR was performed to screen for the presence of three ESBL gene families ( bla CTX‐M bla SHV and bla TEM ) using specific primer (Dallenne et al ).…”

Section: Methodsmentioning

confidence: 99%

Ready‐to‐eat street food: a potential source for dissemination of multidrug‐resistant Escherichia coli epidemic clones in Quito, Ecuador

Zurita

Yánez

Sevillano³

et al. 2020

Lett Appl Microbiol

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Significance and Impact of the Study: This study identified ESBL-producing Escherichia coli epidemic clones: ST131, ST410 and ST744 in ready-to-eat street food samples. Street food is a possible way to spread harm multidrug-resistant (MDR) E. coli strains in the community. Studies to identify the contamination sources of this kind of food are needed to tackle MDR E. coli dissemination. AbstractReady-to-eat food contamination with ESBL-producing Escherichia coli is a growing health concern. Some of these strains also are epidemic clones and can cause community-associated infections that are difficult to treat. In this study, the occurrence of ESBL-producing E. coli contaminated ready-to-eat street food in Quito, Ecuador was evaluated. In total, 150 samples were collected randomly in the most crowded sites of the city. In all, 34 samples (34/150; 22Á6%) were positive for total thermotolerant (44Á5°C) coliforms resistant to cefotaxime. MALDI-TOF analysis identified that the E. coli was found in 20 food samples (20/34; 59%). ESBL gene bla CTX-M-55 was identified in nine isolates, bla CTX-M-15 in six isolates, bla CTX-M-14 in two isolates, and one isolate each harboured bla CTX-M-24 , bla CTX-M-65 , bla CTX-M-55 and bla CTX-M-8 . Phylogenetic groups like A and B1 were the most common, followed by groups D and B2. MLST analysis identified 12 different sequence types (STs), the most common was ST162. Recognized epidemic clonal groups ST410, ST131 and ST744 were encountered. Ready-to-eat street food is a potential way of spreading ESBL-producing E. coli epidemic clones in Quito, Ecuador.Letters in Applied Microbiology ISSN 0266-8254

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Section: Methodsmentioning

confidence: 99%

Ready‐to‐eat street food: a potential source for dissemination of multidrug‐resistant Escherichia coli epidemic clones in Quito, Ecuador

Zurita

Yánez

Sevillano³

et al. 2020

Lett Appl Microbiol

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“…DNA was extracted using the boiling method. 21 The PCRs were performed at a final volume of 25 mL using a green master mix (TSINGKE, China), and the products were visualized by agarose gel (1% w/v) electrophoresis (TSINGKE) stained with ethidium bromide (TSINGKE).…”

Section: Molecular Detection Of Antimicrobial Resistance Genes and Inmentioning

confidence: 99%

A New Phage Cocktail Against Multidrug, ESBL-Producer Isolates of Shigella sonnei and Shigella flexneri with Highly Efficient Bacteriolytic Activity

Shahin

Bouzari

Komijani

et al. 2020

Microbial Drug Resistance

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The globally increasing incidence of antibiotic resistance in pathogenic microorganisms such as Shigella, a cause of human acute gastrointestinal infections, calls for developing effective alternatives. In this study, the antibiotic resistance pattern, extended-spectrum b-lactamase (ESBL)-production, and molecular characteristics of 70 multidrug-resistant isolates belong to the two most frequent species of Shigella genus, that is, Shigella sonnei (44 isolates) and Shigella flexneri (26 isolates) were investigated. These isolates were used to evaluate both specificity and activity of Shigella-specific bacteriophages, vB_SflS-ISF001, vB_SsoS-ISF002, and a cocktail of both. Twelve out of the 21 tested resistance genes were detected in the isolates. About 59% of S. sonnei and 46% of S. flexneri isolates were identified as ESBL producers. The bacteriophages showed a high efficiency of plating (EOP ‡0.5) in about 75% of the isolates. Moreover, the growth of >85% of the isolates was inhibited by the phage cocktail of vB_SflS-ISF001 and vB_SsoS-ISF002. The phage cocktail was effective against a wide range of ESBL-positive and -negative isolates of S. sonnei and S. flexneri. Therefore, this phage cocktail has the potential to inhibit or significantly decrease the spread of drug-resistant Shigella in humans, food chains, and water/wastewater sanitation systems.

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“…Cows in TYLO received once daily SC injection of 10 mg/kg mg tylosin (Tyloject® 20%, Razak Laboratories Co, Iran) for three days, at 21±3 days prior to calving and cows in MARB received single SC injection of 8 mg/kg SC marbofloxacin (Marbox®, Ceva Santé Animale BV, Netherlands) at 21±3 days prior to calving while cows assigned to CONT remained untreated nucA PCR. DNA extraction was performed by rapid boiling according to method by Abdelhai et al (47). Primers were selected on the basis of the published nucleotide sequence of the 613-bp nuc gene (F-CTG GCA TAT GTA TGG CAA TTG TT, R-TAT TGA CCT GAA TCA GCG TTG TCT).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of close up antimicrobial therapies for treatment and prevention of subclinical mastitis in the herds with high prevalence of Staphylococcus aureus

Amiri

Rad

Heidarpour

et al. 2019

Preprint

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Background: This field trial was conducted to describe and compare the efficacy of two treatment regimens comprising of pre-calving systemic administrations of marbofloxacin and tylosin in combination with dry cow therapy. For more precise detection of Staphylococcus aureus IMI, PCR (nucA gene) and RAPD-PCR genotyping have been applied. A total of 841 quarters from 212 dry cows within two herds were assigned to three groups TYLO, MARB and CONT. At 21±3 days prior to parturition, tylosin was injected for three days (10 mg/kg, SC) to cows from TYLO and at the same time, single injection of marbofloxacin (8 mg/kg, SC) was administered to cows from MARB. Cows assigned to CONT served as an untreated control. All quarters allocated to the groups received intra-mammary infusion of Kanaclox DC® at drying off. Milk samples from all quarters at one week, before last milking of dry off, 3 and 7 DIM were obtained for bacteriological and oxidative analyses. Results: Despite the fact that no significant differences in total cure rate within the groups was demonstrated, the S. aureus cure rate achieved in TYLO and MARB were 74 and 73.5% respectively both of which being significantly higher than in CONT (58.1%). No significant differences in total new IMI were observed in TYLO (40.9%) and MARB (38.5%) groups compared with CONT group (42.5%). Furthermore the rate of new S. aureus IMI was higher in TYLO (24.5%) and MARB (24.1%) than in CONT (13.8%) group. Clinical mastitis rate in cows that received systemic injections (TYLO=3.8%, MARB=5.8%) was significantly lower than those detected in control cows (CONT=11.3%). Paired S. aureus isolates from dry off and post-calving have been clustered into 9 different RAPD types (A-I). 8 paired strains collected at dry off were identical to those at post-calving and 35 strains had more than 60% of dissimilarity. Conclusion: The study demonstrates that dealing with herds in which S. aureus IMIs are prevalent, using close up broad-spectrum systemic antimicrobial agents can be unrewarding for preventing new S. aureus IMIs. However close up administration of systemic antimicrobials have the potential to reduce the incidence of clinical mastitis during lactation.

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Comparative Study of Rapid DNA Extraction Methods of Pathogenic Bacteria

Cited by 23 publications

References 23 publications

Ready‐to‐eat street food: a potential source for dissemination of multidrug‐resistant Escherichia coli epidemic clones in Quito, Ecuador

Ready‐to‐eat street food: a potential source for dissemination of multidrug‐resistant Escherichia coli epidemic clones in Quito, Ecuador

A New Phage Cocktail Against Multidrug, ESBL-Producer Isolates of Shigella sonnei and Shigella flexneri with Highly Efficient Bacteriolytic Activity

Evaluation of close up antimicrobial therapies for treatment and prevention of subclinical mastitis in the herds with high prevalence of Staphylococcus aureus

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