scholarly journals Molecular Typing of Staphylococcus Aureus Isolated from Various Environmental Sources

2020 ◽  
Vol 5 (6) ◽  
pp. 1243-1248
Author(s):  
Ingilala Satheesh Kumar ◽  
Nadakuditi Venkata Raju ◽  
Hanumohan R. Konatham
Keyword(s):  
Molecular Typing ◽  
Dna Isolation ◽  
Profile Analysis ◽  
Epidemiological Studies ◽  
Genomic Dna Isolation ◽  
Rapd Pcr ◽  
Pure Cultures ◽  
Host Origin ◽  
Different Sources

RAPD-PCR was employed for the characterization of S. aureus isolates from different sources such as soil, water, milk, meat and skin swab etc.,. Isolated pure cultures of S.aureus strains were subjected to genomic DNA isolation, purification, separation and quantification. Isolated DNA samples were distinguished by using 4 different random primers. Genome profile analysis obtained from the S.aureus demonstrated that it was possible to differentiate the S. aureus strains from different sources by RAPD technique. Results indicate possible relationships between host origin and genetic variation among S. aureus isolates from various sources. This RAPD method was thus useful for epidemiological studies of the S. aureus flora.

scholarly journals Assessment of the Genetic Diversity among Arcobacters Isolated from Poultry Products by Using Two PCR-Based Typing Methods

2002 ◽  
Vol 68 (5) ◽  
pp. 2172-2178 ◽  
Author(s):  
Kurt Houf ◽  
Lieven De Zutter ◽  
Jan Van Hoof ◽  
Peter Vandamme
Keyword(s):  
Genotypic Diversity ◽  
Epidemiological Studies ◽  
Dna Templates ◽  
Poultry Products ◽  
Large Numbers ◽  
Rapd Pcr ◽  
Arcobacter Butzleri ◽  
Typing Methods ◽  
Dna Template

ABSTRACT In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and randomly amplified polymorphic DNA PCR (RAPD-PCR) were optimized for characterization of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. In addition, a simple and rapid DNA extraction method was tested for use in both typing procedures. Both methods had satisfactory typeability and discriminatory power, but the fingerprints generated with ERIC-PCR were more reproducible and complex than those obtained with RAPD-PCR. The use of nondiluted boiled cell suspensions as DNA templates was found to be very useful in ERIC-PCR. Characterization of large numbers of Arcobacter isolates is therefore preferably performed by the ERIC-PCR procedure. Isolates for which almost identical ERIC fingerprints are generated may subsequently be characterized by RAPD-PCR, although adjustment and standardization of the amount of the DNA template are necessary. In the second part of this study, the genotypic diversity of arcobacters present on broiler carcasses was assessed by using both typing methods. A total of 228 cultures from 24 samples were examined after direct isolation and enrichment. The isolates were identified by using a multiplex PCR as A. butzleri (n = 182) and A. cryaerophilus (n = 46). A total of 131 types (91 A. butzleri types and 40 A. cryaerophilus types) were discerned without discordance between the two typing techniques. The analysis of the poultry isolates showed that poultry products may harbor not only more than one species but also multiple genotypes. All genotypes were confined to one poultry sample, and only three genotypes were found after simultaneous enrichment and direct isolation. These results demonstrate that different outcomes can be obtained in epidemiological studies depending on the isolation procedure used and the number of isolates characterized.

scholarly journals MOLECULAR CHARACTERIZATION OF WHEAT GENOTYPES (TRITICUM AESTIVUM L.)

2017 ◽  
Vol 8 ◽  
pp. 29-36
Author(s):  
SHARAD PAWAR, MUNNESHKUMAR, V. M. JAMBHALE
Keyword(s):  
Dna Isolation ◽  
Pcr Amplification ◽  
Triticum Aestivum L ◽  
Dice Similarity Coefficient ◽  
Wheat Genotypes ◽  
Genomic Dna Isolation ◽  
Ssr Primers ◽  
Minimum Number ◽  
Maximum Similarity

Molecular diversity in thirty wheat genotypes was done. For this the genomic DNA isolation was carried out and which were then subjected to PCR amplification using twenty SSR primers. Out of these twenty SSR primers, eighteen yielded amplifications and showed polymorphism. Total 93 loci were generated by amplification with 18 polymorphic primers, all of which 93 loci were polymorphic i.e. 100%. Among the SSR primers, BARC-170, WMC-44, produced maximum number of 2 loci. The size of amplification products ranged from 102 bp to 805 bp. All SSR primers showed 100 % polymorphism and all primers had more than 0.50 PIC value except one primer. Maximum PIC value 0.17 was observed in WMC-468. The maximum number of bands were observed in NIAW-2721 (28 bands), whereas minimum number of bands were present in NIAW-301 and NIAW-2539 (19 bands). The dice similarity coefficient values ranged from 0.50 to 0.95. Maximum similarity value of 0.95 was noticed between NIAW-2891 and NIAW-2837, while minimum similarity value of 0.50 was observed among NIAW-2595, NIAW-2874, NIAW-2995 and NIAW-2725. The consensus tree software revealed two major clusters.

scholarly journals Molecular typing of Staphylococcus aureus from different sources by RAPD-PCR analysis

Heliyon ◽  
2019 ◽  
Vol 5 (8) ◽  
pp. e02231 ◽  
Author(s):  
Sahar Zare ◽  
Abdollah Derakhshandeh ◽  
Masoud Haghkhah ◽  
Zahra Naziri ◽  
Azar Motamedi Broujeni
Keyword(s):  
Staphylococcus Aureus ◽  
Molecular Typing ◽  
Pcr Analysis ◽  
Rapd Pcr ◽  
Different Sources

Applications of SIMS to electronic materials and devices

1992 ◽  
Vol 50 (2) ◽  
pp. 1682-1683
Author(s):  
S.F. Corcoran
Keyword(s):  
Depth Distribution ◽  
Secondary Ion Mass Spectrometry ◽  
Semiconductor Device ◽  
Electronic Materials ◽  
Profile Analysis ◽  
Semiconductor Industry ◽  
Small Diameter ◽  
Depth Resolution ◽  
Detection Sensitivity

Over the past decade secondary ion mass spectrometry (SIMS) has played an increasingly important role in the characterization of electronic materials and devices. The ability of SIMS to provide part per million detection sensitivity for most elements while maintaining excellent depth resolution has made this technique indispensable in the semiconductor industry. Today SIMS is used extensively in the characterization of dopant profiles, thin film analysis, and trace analysis in bulk materials. The SIMS technique also lends itself to 2-D and 3-D imaging via either the use of stigmatic ion optics or small diameter primary beams.By far the most common application of SIMS is the determination of the depth distribution of dopants (B, As, P) intentionally introduced into semiconductor materials via ion implantation or epitaxial growth. Such measurements are critical since the dopant concentration and depth distribution can seriously affect the performance of a semiconductor device. In a typical depth profile analysis, keV ion sputtering is used to remove successive layers the sample.

An Outbreak of ESBL-producing Klebsiella pneumoniae in an Iranian Referral Hospital: Epidemiology and Molecular Typing

2019 ◽  
Vol 19 (1) ◽  
pp. 46-54 ◽  
Author(s):  
Shima Mahmoudi ◽  
Babak Pourakbari ◽  
Aliakbar Rahbarimanesh ◽  
Mohammad Reza Abdosalehi ◽  
Keyghobad Ghadiri ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Nosocomial Infections ◽  
Molecular Typing ◽  
Genetic Relatedness ◽  
Referral Hospital ◽  
Limited Information ◽  
Hospital Epidemiology ◽  
Rapd Pcr ◽  
Published Report ◽  
Random Amplification

Introduction: Klebsiella pneumoniae is a common cause of nosocomial infections; however, there is limited information in Iran regarding nosocomial outbreaks due to extended-spectrum β–lactamase (ESBL) producing K pneumoniae strains, particularly using molecular methods. The present study focused on the molecular mechanism of ESBL resistance and genetic relatedness in K. pneumoniae isolates causing nosocomial infections in an Iranian referral hospital. Material and Methods: This study evaluated the antimicrobial resistance and molecular epidemiology of K. pneumoniae causing nosocomial infections in children between October 2013 and March 2014. The ESBL detection was carried out for all the isolates by the CLSI method and PCR was carried out for the detection of the blaSHV, blaTEM, and blaCTX-M genes among ESBL-producing K. pneumonia. Molecular typing of the K. pneumoniae was performed using random amplification of polymorphic DNA-polymerase chain reaction (RAPD-PCR). Results: A total of 30 isolates of K. pneumoniae were used for epidemiological analysis. High rates of resistance to cefotaxime (n=29, 97%), cefazolin (n=29, 97%), cefepime (n=25, 83%) and gentamicin (n=23, 77%) were observed. A total of 29 strains (97%) produced ESBLs. The frequency of blaSHV, blaCTX-M and blaTEM genes among these isolates was 83% (n=25), 70% (n=21) and 57% (n=17), respectively. Surprisingly 11 isolated (37%) carried blaSHV, blaCTX-M and blaTEM genes simultaneously. Moreover, the concurrent presence of “blaSHV and blaCTX-M” and “blaSHV and blaTEM” was seen in 8 (27%) and 4 (13%) isolates, respectively. RAPDPCR analyses revealed that K. pneumoniae isolates belonged to 2 RAPD-PCR types among which one cluster counted for 28 isolates. Conclusion: To our knowledge, this is the first published report of a nosocomial outbreak of ESBL-producing K. pneumoniae in children in Iran. Although the epidemiology of nosocomial infections with ESBL-producing organisms has not yet been explored in depth in Iran, our findings suggest that ESBL-producing organisms are already an established public health threat in our country.

scholarly journals Microfluidic Tools for Enhanced Characterization of Therapeutic Stem Cells and Prediction of Their Potential Antimicrobial Secretome

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 750
Author(s):  
Pasquale Marrazzo ◽  
Valeria Pizzuti ◽  
Silvia Zia ◽  
Azzurra Sargenti ◽  
Daniele Gazzola ◽  
...  
Keyword(s):  
Stem Cells ◽  
Defense Mechanisms ◽  
Live Cells ◽  
Label Free ◽  
Bacterial Diseases ◽  
Prospective Application ◽  
Therapy Strategies ◽  
Stromal Stem Cells ◽  
Different Sources

Antibiotic resistance is creating enormous attention on the development of new antibiotic-free therapy strategies for bacterial diseases. Mesenchymal stromal stem cells (MSCs) are the most promising candidates in current clinical trials and included in several cell-therapy protocols. Together with the well-known immunomodulatory and regenerative potential of the MSC secretome, these cells have shown direct and indirect anti-bacterial effects. However, the low reproducibility and standardization of MSCs from different sources are the current limitations prior to the purification of cell-free secreted antimicrobial peptides and exosomes. In order to improve MSC characterization, novel label-free functional tests, evaluating the biophysical properties of the cells, will be advantageous for their cell profiling, population sorting, and quality control. We discuss the potential of emerging microfluidic technologies providing new insights into density, shape, and size of live cells, starting from heterogeneous or 3D cultured samples. The prospective application of these technologies to studying MSC populations may contribute to developing new biopharmaceutical strategies with a view to naturally overcoming bacterial defense mechanisms.

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