scholarly journals mecA-Gene Detection and MDR Profile of S. Aureus Isolates From Patients Attending The Referral Hospitals of Amhara Reginal State, Ethiopia

2021 ◽  
Author(s):  
Feleke Moges ◽  
Tadele Tamiru ◽  
Azanaw Amare ◽  
Getachew Mengistu ◽  
Wondwossen Abebe ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Meca Gene ◽  
Surrogate Marker ◽  
Pcr Amplification ◽  
Resistance Rate ◽  
Molecular Techniques ◽  
High Rate ◽  
Amhara Region ◽  
Genomic Dna Isolation ◽  
Referral Hospitals

Abstract Background: Staphylococcus aureus causes different types of human infections, and has an ability to develop resistance to many antibiotics. There is scarcity of data on mecA gene and MDR profiles of this organism in developing countries, like Ethiopia. The aim of the present study is therefore, to investigate MDR profiles and mecA-gene profile of S. aureus from Referral Hospitals of Amhara Reginal State. Methods: Of the total of 110 isolates collected from Amhara Region Referral Hospitals, 70 MDR isolates were further processed for isolation of S. aureus mecA gene. Genomic DNA was isolated using SIGMA ALDRICH genomic DNA isolation Kit for Gram positive bacteria. Amplification of S. aureus mecA gene was done using amplicon size of 533 bp. Agarose gel electrophoresis was prepared with 1.5% agarose by TAE solvent and 0.5μg/mL of ethidium bromide. Electrophoresis was carried out for 1 hour and a half (7 Volts/ cm²) in 1X tris acetate EDTA (TAE) buffer. Data were analysed by using descriptive statistics.Results: Majority of the isolates were recovered from patients age less than 5 years, 51 (36.7%) and least from age greater than 60 years, 6 (4.3%). Most of the isolates were from blood, 61 (43.9%), followed by wound, 45 (32.4%). High resistance rate was observed in penicillin 81 (73.6%), followed by cotrimoxazole 78 (70.9%), ceftriaxone 76 (69%), erythromycin 66 (60%) and tetracycline 65 (59.1%). Phenotypically, considering cefoxitin as a surrogate marker, 38 (34.5%) of the isolates were methicillin resistant. The overall MDR isolates were 92 (83.6%). The PCR amplification result of mecA gene was 14 (20%).Conclusions and recommendations: There are high rate of MDR and MRSA isolates of S. aureus. PCR amplification result indicates 20% of MRSA isolates are mecA gene producers. Largescale study for detection of MDR strains of S. aureus including MRSA using molecular techniques should be encouraged in Amhara region.

Investigations of Genetic Variation Between Olive (Olea europaea L.) Cultivars Using Arbitrarily Primed Polymerase Chain Reaction (AP-PCR)

2003 ◽  
Vol 58 (11-12) ◽  
pp. 837-842 ◽  
Author(s):  
Feray Kockar ◽  
Rahsan Ilıkcı
Keyword(s):  
Genetic Relationship ◽  
Genomic Dna ◽  
Pcr Amplification ◽  
Dna Fingerprint ◽  
Factors Affecting ◽  
Genomic Dna Isolation ◽  
Amplification Protocol ◽  
Olive Cultivars ◽  
Arbitrarily Primed Pcr ◽  
Relationship Of

Abstract Characterization and selection of olive clones for the production of olive oil is essential in Turkey because of its profitable exploitation. AP-PCR (Arbitrarily-Primed PCR) is a technique that can distinguish the genetic relationship among plant species and other organisms. In this study, AP-PCR approach was used in order to determine the genetic relationship of different six olive clones. The purity of DNA is one of the most important factors affecting the product of the AP-PCR method. In this respect, modified genomic DNA isolation procedure from Oleae europaea clones was developed so that this procedure can be used to obtain plant genomic DNA from diverse aromatic plants, which produce essential oils and secondary metabolites. By following the optimized AP-PCR amplification protocol, unique DNA fingerprint profiles for each olive clone were produced. AP-PCR-generated unique DNA fingerprint profiles can be used in the identification, distribution and diversity of various olive cultivars.

scholarly journals Optimizing the pure genomic DNA isolation procedure for Plectranthus amboinicus DNA – A prerequisite for further genomic Studies

2017 ◽  
Vol 2 (4) ◽  
Author(s):  
S. Manikandan ◽  
S. Ansarali ◽  
G.M. Alagu Lakshmanan
Keyword(s):  
Dna Barcoding ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Leaf Tissue ◽  
Isolation Procedure ◽  
Modified Method ◽  
Genomic Dna Isolation ◽  
Young Leaves ◽  
Plectranthus Amboinicus

The DNA isolation procedure for different plant groups have been studied and standardized.  The isolation of pure genomic DNA is the most essential component for any type of molecular studies.  The present work is aimed to identify suitable DNA markers for the amplification of P.amboinicus  DNA  becomes a great hurdle  for DNA barcoding studies carried out by rbcL and matK primers Used in the members of Lamiaceae. To solve this problem, The DNA was extracted by three methods from fresh young leaf tissue of P.amboinicus. After the evaluationthe outcome of these methods, one most suitable modified method was selected for isolating DNA from young leaves of P.amboinicus and selected for suitable DNA barcoding markers for PCR amplification. The quality and quantity of DNAs are a prerequisite for genetic studies for a variety of plants including P.amboinicus. The quantity and quality of the DNA extracted by this method wasused for suitable DNA barcoding markers selection.

scholarly journals Optimizing Genomic DNA Isolation and PCR Amplification For Pasak Bumi (Eurycoma longifolia)

2019 ◽  
Vol 1 (2) ◽  
pp. 30
Author(s):  
Arida Susilowati ◽  
Henti Hendalastuti Rachmat ◽  
Ahmad Baiquni Rangkuti ◽  
Deni Elfiati ◽  
Ami Ambarwati
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Eurycoma Longifolia ◽  
Genomic Dna Isolation

.

scholarly journals Comparative Analysis of the Genomic DNA Isolation Methods on Inula sp. (Asteraceae)

2016 ◽  
Vol 8 (4) ◽  
pp. 451-455
Author(s):  
Emre SEVİNDİK ◽  
Fatih COŞKUN ◽  
Zehra Tuğba MURATHAN ◽  
Mehmet Yavuz PAKSOY ◽  
Veysel UZUN
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Low Cost ◽  
Pcr Amplification ◽  
Isoamyl Alcohol ◽  
Dna Amount ◽  
Genomic Dna Isolation ◽  
Asteraceae Family ◽  
Positive Results ◽  
Commercial Kit

Simple, fast, low-cost and high throughput protocols are required for DNA isolation of plant species. In this study, phenol chloroform isoamyl alcohol and commercial (Sigma) DNA isolation kit methods were applied on some Inula species that belong to Asteraceae family. Genomic DNA amounts, A260, A280, A260/A230 and purity degrees (A260/A280) that were obtained through both methods were measured through electrophoresis and spectrophotometer. Additionally, PCR amplification was realized by primer pairs specific to nrDNA ITS, cpDNA ndhF (972F-1603R) and trnL-F regions. Results showed that maximum genomic DNA in nanograms obtained by phenol chloroform isoamyl alcohol method. The study also revealed that I. macrocephala had the maximum DNA and I. heterolepis had the minimum DNA amount. A260/A280 purity degrees showed that the highest and lowest purity in gDNAs obtained through phenol-choloform isoamyl alcohol method were in I.aucheriana and I. salicina, respectively. The highest and lowest purity degrees of gDNAs obtained through commercial kit was observed in I. fragilis and I. macrocephala samples, respectively. PCR amplification results showed that while band profiles of each three regions (ITS, trnL-F and ndhF) did not yield positive results in PCR amplifications using phenol-choloform isoamyl alcohol method; PCR band profiles obtained through commercial kit yielded positive results. As a result, it is fair to say that the relation of genomic DNA with PCR was found to be more efficient although the maximum amount of genomic DNA was obtained through phenol chloroform isoamyl alcohol method. 

scholarly journals Development of a Competent and Trouble Free DNA Isolation Protocol for Downstream Genetic Analyses in Glycine Species

2016 ◽  
Vol 4 (8) ◽  
pp. 700 ◽  
Author(s):  
Muhammad Amjad Nawaz ◽  
Faheem Shehzad Baloch ◽  
Hafiz Mamoon Rehman ◽  
Bao Le ◽  
Fahima Akther ◽  
...  
Keyword(s):  
Molecular Biology ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Plant Molecular Biology ◽  
Pcr Amplification ◽  
Cost Effective ◽  
Extraction Methods ◽  
Glycine Species ◽  
Genomic Dna Isolation

Extraction of deoxyribose nucleic acid (DNA) from plants is preliminary step in molecular biology. Fast and cost effective genomic DNA isolation from Glycine species for downstream application is a major bottleneck. Here we report a high throughput and trouble free method for genomic DNA extraction from leaf and seeds of Glycine species with high quality and quantity. Protocol reports the optimization by employing different concentrations of CTAB and PVP in extraction buffer. Efficiency of optimized protocol was compared with frequently used DNA extraction methods. Wide adoptability and utility of this protocol was confirmed by DNA extraction from leaves as well as seeds of G. max, G. soja, G. tomentella and G. latifolia. Extracted DNA was successfully subjected to PCR amplification of five microsatellite markers and four putative glycosyltransferase genes. DNA extraction protocol is reproducible, trouble free, rapid and can be adopted for plant molecular biology applications.

Staphylococcus aureus nasal colonization among Vietnamese adults: prevalence, risk factors and antibiotic susceptibility profile

MedPharmRes ◽  
2018 ◽  
Vol 2 (2) ◽  
pp. 21-31
Author(s):  
Nguyen Phan ◽  
Hien Pham ◽  
Thuc Nguyen ◽  
Hoai Nguyen
Keyword(s):  
Risk Factors ◽  
Staphylococcus Aureus ◽  
Antibiotic Susceptibility ◽  
Nasal Carriage ◽  
Resistance Rate ◽  
High Rate ◽  
Weight History ◽  
Potential Risk Factors ◽  
History Of ◽  
Vietnamese Community

Staphylococcus aureus (S. aureus) has long been recognized as an important human pathogen causing many severe diseases. It is also a part of human normal flora with its ecological niche in the human anterior nares. This study focused on screening S. aureus nasal carriage in community and its relationship to human physiological and pathological factors which have not been studied in Vietnam previously. Two hundred and five volunteers in Ho Chi Minh City from 18 to 35 and over 59 years old both male and female participated in the study. Result showed that the prevalence of S. aureus nasal carriage in southern Vietnamese community was relatively low, only 11.2% (23/205), much lower than that in other international reports on human S. aureus. In addition, nasal carriage of the older age group (> 59 years old, 13.7%) was higher than that of younger age (18-35 years old, 10.4%). Other potential risk factors such as gender, career, height, weight, history of antibiotic usage, daily nasal wash, use of nasal medication sprays, acne problems, smoking and nasal problems showed no significant impact on S. aureus carriage. The obtained S. aureus nasal isolates were all sensitive to vancomycin. Lincomycin and tetracycline had low resistance rate with 4.3 % and 17.4 %, respectively. However, the isolates showed particularly high rate of multidrug resistance (54.2%) In summary, our data provided researchers an overview on S. aureus nasal carriage and antibiotic susceptibility profile of the community- isolated S. aureus in Vietnam. This would serve as valuable information on assessing risk of community-acquired S. aureus infections.

scholarly journals Single and Double Infections with Wolbachia in the Parasitic Wasp Nasonia vitripennis Effects on Compatibility

Genetics ◽  
1996 ◽  
Vol 143 (2) ◽  
pp. 961-972 ◽  
Author(s):  
Marie-Jeanne Perrot-Minnot ◽  
Li Rong Guo ◽  
John H Werren
Keyword(s):  
Genomic Dna ◽  
Rapid Development ◽  
Parasitic Wasp ◽  
Pcr Amplification ◽  
Bacterial Strains ◽  
Nasonia Vitripennis ◽  
Larval Diapause ◽  
Wide Range ◽  
Reproductive Incompatibility ◽  
Infected Line

Abstract Wolbachia are cytoplasmically inherited bacteria responsible for reproductive incompatibility in a wide range of insects. There has been little exploration, however, of within species Wolbachia polymorphisms and their effects on compatibility. Here we show that some strains of the parasitic wasp Nasonia vitripennis are infected with two distinct bacterial strains (A and B) whereas others are singly infected (A or B). Double and single infections are confirmed by both PCR amplification and Southern analysis of genomic DNA. Furthermore, it is shown that prolonged larval diapause (the overwintering stage of the wasp) of a double-infected strain can lead to stochastic loss of one or both bacterial strains. After diapause of a double-infected line, sublines were produced with AB, A only, B only or no Wolbachia. A and B sublines are bidirectionally incompatible, whereas males from AB lines are unidirectionally incompatible with females of A and B sublines. Results therefore show rapid development of bidirectional incompatibility within a species due to segregation of associated symbiotic bacteria.

scholarly journals Genetic Diversity of Salvia Species Assessed by ISSR and RAPD Markers

2016 ◽  
Vol 44 (2) ◽  
pp. 431-436 ◽  
Author(s):  
Masoumeh YOUSEFIAZARKHANIAN ◽  
Ali ASGHARI ◽  
Jafar AHMADI ◽  
Behvar ASGHARI ◽  
Ali Ashraf JAFARI
Keyword(s):  
Genetic Diversity ◽  
Genetic Polymorphisms ◽  
Genomic Dna ◽  
Rapd Markers ◽  
Genetic Relationships ◽  
Genetic Variations ◽  
Molecular Techniques ◽  
Wild Populations ◽  
Similarity Coefficients ◽  
The World

The genus Salvia includes an enormous assemblage of nearly 1,000 species dispersed around the world. Due to possible threats to this genus, there is an immediate requirement to evaluate the diversity of its wild populations. ISSR and RAPD molecular techniques were used to evaluate the genetic relationships among twenty-one ecotypes of eight Salvia species. Amplification of genomic DNA using 23 primers (15 RAPD and eight ISSR) produced 280 bands, of which 91% were polymorphic. The results of marker parameters showed no clear difference between two marker systems. It was generally observed that both ISSR and RAPD markers had similar efficiency in detecting genetic polymorphisms with remarkable ability to differentiate the closely related ecotypes of Salvia. Nei’s similarity coefficients for these techniques ranged from 0.48 to 0.98. Based on the results of clustering, PCoA and AMOVA, the genetic diversity between and within species was confirmed. So, conservation and domestication of the genus Salvia must be due to levels of genetic variations.

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