scholarly journals RAPD-PCR as a Molecular Discriminative Technique for Human Pathogenic Bacteria - A Review

2015 ◽  
Vol 42 ◽  
pp. 13-17
Author(s):  
Partha Pal
Keyword(s):  
Pathogenic Bacteria ◽  
Present Report ◽  
Random Amplified Polymorphic Dna ◽  
Viable Cell ◽  
Immunological Methods ◽  
Species Differentiation ◽  
Bacterial Isolates ◽  
Rapd Pcr ◽  
Microbiological Methods ◽  
Biological Technique

The application of random amplified polymorphic DNA- polymerase chain reaction (RAPD-PCR) was found to be a simple, cheap and rapid tool to discriminate human pathogenic bacterial isolates especially at intraspecific level. This molecular biological technique relies on the use of random oligonucleotide primers that arbitrarily amplifies specific regions of the genome which gives rise to a unique genomic fingerprint of the strains under investigations. With continued development of novel molecular-based technologies for rapid, high-throughput detection of food borne pathogenic bacteria, the future of conventional microbiological methods such as viable cell enumeration, selective isolation of bacteria on commercial media, and immunoassays seems tenuous. Approaches that enhance recovery of sub lethally injured bacteria, differentiation among species, differentiation among bacteria of interest using biochemical profiling, enumeration using impedance technology, techniques to confirm the presence of target pathogens using immunological methods, and bioluminescence applications for hygiene monitoring are of utmost need in identifying and combating the human pathogenic isolates. The aim of this study is to esti­mate the efficiency of RAPD-PCR technique in assessing the genetic diversity of diseases causing bacterial isolates. The use of RAPD-PCR in evaluating the genomic variability among the pathogenic strains belonging to different genus are also been discussed in the present report.

A review of conventional detection and enumeration methods for pathogenic bacteria in food

2004 ◽  
Vol 50 (11) ◽  
pp. 883-890 ◽  
Author(s):  
Kiev S Gracias ◽  
John L McKillip
Keyword(s):  
Pathogenic Bacteria ◽  
Pathogen Detection ◽  
Low Cost ◽  
Viable Cell ◽  
Ease Of Use ◽  
Food Microbiology ◽  
Immunological Methods ◽  
Commercial Media ◽  
Microbiological Methods ◽  
High Throughput Detection

With continued development of novel molecular-based technologies for rapid, high-throughput detection of foodborne pathogenic bacteria, the future of conventional microbiological methods such as viable cell enumeration, selective isolation of bacteria on commercial media, and immunoassays seems tenuous. In fact, a number of unique approaches and variations on existing techniques are currently on the market or are being implemented that offer ease of use, reliability, and low cost compared with molecular tools. Approaches that enhance recovery of sublethally injured bacteria, differentiation among species using fluorogenics or chromogenics, dry plate culturing, differentiation among bacteria of interest using biochemical profiling, enumeration using impedence technology, techniques to confirm the presence of target pathogens using immunological methods, and bioluminescence applications for hygiene monitoring are summarized here and discussed in relation to their specific advantages or disadvantages when implemented in a food microbiology setting.Key words: food pathogen, detection, enumeration methods, food safety.

Molecular identification and typing of lactobacilli isolated from kefir grains

2005 ◽  
Vol 73 (1) ◽  
pp. 20-27 ◽  
Author(s):  
Lucrecia Delfederico ◽  
Axel Hollmann ◽  
Mariano Martínez ◽  
N. Gabriel Iglesias ◽  
Graciela De Antoni ◽  
...  
Keyword(s):  
Restriction Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Restriction Enzymes ◽  
23S Rrna ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Bacterial Isolates ◽  
Good Tool ◽  
Rapd Pcr ◽  
Kefir Grains

Seventeen heterofermentative lactobacilli isolated from kefir grains were characterized by molecular methods. Bacterial isolates were identified by amplification of 16S rRNA gene and analysis by Amplified Ribosomal DNA Restriction Analysis (ARDRA), using the restriction enzymes Hae III, Dde I, and Hinf I. ARDRA analysis of lactobacilli isolates showed, for each enzyme used, a same banding pattern between the heterofermentative lactobacilli and the reference strains Lactobacillus kefir JCM 5818 and Lb. kefir ATCC 8007. Other reference lactobacilli and one homofermentative isolate showed differences in at least one of these patterns. The 16S-23S rRNA spacer region was also used to discriminate the bacterial isolates at the species level. The data obtained from the analysis of spacer region confirmed that sequencing of this genome region is a good tool for a reliable identification of members of Lb. kefir species. Genotyping of isolates was performed by Random Amplified Polymorphic DNA (RAPD-PCR) analysis using M13, Coc, ERIC-2 and 1254 primers. Patterns obtained allowed the differentiation of isolates in three groups. The three clusters showed by RAPD-PCR analysis could be correlated with at least three different strains of Lb. kefir species in the group of heterofermentative lactobacilli isolates obtained from Argentinian kefir grains.

scholarly journals Oenological Potential of Autochthonous Saccharomyces cerevisiae Yeast Strains from the Greek Varieties of Agiorgitiko and Moschofilero

Beverages ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 27
Author(s):  
Dimitrios Kontogiannatos ◽  
Vicky Troianou ◽  
Maria Dimopoulou ◽  
Polydefkis Hatzopoulos ◽  
Yorgos Kotseridis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Alcoholic Fermentation ◽  
Ethanol Tolerance ◽  
Random Amplified Polymorphic Dna ◽  
Yeast Strains ◽  
Rapd Pcr ◽  
Autochthonous Yeast ◽  
Polymerase Chain ◽  
Grape Varieties ◽  
H2s Production

Nemea and Mantinia are famous wine regions in Greece known for two indigenous grape varieties, Agiorgitiko and Moschofilero, which produce high quality PDO wines. In the present study, indigenous Saccharomyces cerevisiae yeast strains were isolated and identified from spontaneous alcoholic fermentation of Agiorgitiko and Moschofilero musts in order to evaluate their oenological potential. Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) recovered the presence of five distinct profiles from a total of 430 yeast isolates. The five obtained strains were evaluated at microvinifications trials and tested for basic oenological and biochemical parameters including sulphur dioxide and ethanol tolerance as well as H2S production in sterile grape must. The selected autochthonous yeast strains named, Soi2 (Agiorgitiko wine) and L2M (Moschofilero wine), were evaluated also in industrial (4000L) fermentations to assess their sensorial and oenological characteristics. The volatile compounds of the produced wines were determined by GC-FID. Our results demonstrated the feasibility of using Soi2 and L2M strains in industrial fermentations for Agiorgitiko and Moschofilero grape musts, respectively.

scholarly journals Survival Strategy of Erwinia amylovora against Copper: Induction of the Viable-but-Nonculturable State

2006 ◽  
Vol 72 (5) ◽  
pp. 3482-3488 ◽  
Author(s):  
M�nica Ordax ◽  
Ester Marco-Noales ◽  
Mar�a M. L�pez ◽  
Elena G. Biosca
Keyword(s):  
Pathogenic Bacteria ◽  
Erwinia Amylovora ◽  
Control Plant ◽  
Viable Cell ◽  
Epifluorescence Microscopy ◽  
Survival Strategy ◽  
Vbnc State ◽  
Tetrazolium Chloride ◽  
Viable But Nonculturable ◽  
Cell Counts

ABSTRACT Copper compounds, widely used to control plant-pathogenic bacteria, have traditionally been employed against fire blight, caused by Erwinia amylovora. However, recent studies have shown that some phytopathogenic bacteria enter into the viable-but-nonculturable (VBNC) state in the presence of copper. To determine whether copper kills E. amylovora or induces the VBNC state, a mineral medium without copper or supplemented with 0.005, 0.01, or 0.05 mM Cu2+ was inoculated with 107 CFU/ml of this bacterium and monitored over 9 months. Total and viable cell counts were determined by epifluorescence microscopy using the LIVE/DEAD kit and by flow cytometry with 5-cyano-2,3-ditolyl tetrazolium chloride and SYTO 13. Culturable cells were counted on King's B nonselective solid medium. Changes in the bacterial morphology in the presence of copper were observed by scanning electron microscopy. E. amylovora entered into the VBNC state at all three copper concentrations assayed, much faster when the copper concentration increased. The addition of different agents which complex copper allowed the resuscitation (restoration of culturability) of copper-induced VBNC cells. Finally, copper-induced VBNC cells were virulent only for the first 5 days, while resuscitated cells always regained their pathogenicity on immature fruits over 9 months. These results have shown, for the first time, the induction of the VBNC state in E. amylovora as a survival strategy against copper.

scholarly journals Detection of pathogenic bacteria associated with earphones used by students of Stamford University Bangladesh

2020 ◽  
Vol 10 (1) ◽  
pp. 1-4
Author(s):  
Omor Ahmed Chowdhury ◽  
Md Raihan Ahmed ◽  
Md Raihan Dipu ◽  
Md Aftab Uddin
Keyword(s):  
Pathogenic Bacteria ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
Disk Diffusion Method ◽  
Antibiotic Resistant ◽  
Potential Source ◽  
Different Types ◽  
Staphylococcus Spp

The use of earphones has increased in recent times throughout the world especially among the different level of students such as school, college or university who have a higher tendency of sharing these among them. Unlike airline headsets, headphones and stethoscope ear-pieces, ear phones are often shared by multiple users and can be a potential medium for transmission of pathogens, which can give rise to various ear related infections. The objective of this study was to detect the pathogenic bacteria from the ear-phones used by the students of Stamford University Bangladesh. A total of 16 ear-phone swabs were collected by sterile cotton swabs. The swabs were inoculated onto blood agar and incubated aerobically overnight at 37oC. Microscopic observation and standard biochemical tests were performed to confirm the identification of all the bacterial isolates. Six presumptively identified Staphylococcus spp. (38%) were tested against six different types of antibiotics following Kirby-Bauer disk diffusion method. Isolates were found to be 84% resistant against Cotrimoxazole and demonstrated 100% sensitivity to Vancomycin and Ciprorofloxacin. The findings of this study suggest the users to disinfect their respective ear phones and not to exchange them as they may act as a potential source to transfer pathogenic and antibiotic resistant bacteria among the ear phone users. Stamford Journal of Microbiology, Vol.10 (1) 2020: 1-4

scholarly journals Karakteristik Genus Bakteri Pada Karkas Ayam Broiler Dari Swalayan di Kota Pontianak

2018 ◽  
Vol 7 (3) ◽  
Author(s):  
Prianti Rahmawati Diah Wulandari Rousdy
Keyword(s):  
Broiler Chicken ◽  
Pathogenic Bacteria ◽  
Chicken Meat ◽  
Bacterial Isolates ◽  
Bacterial Genus ◽  
Chicken Carcass

The availability  of nutrients in chicken carcasses can cause chicken meat to be an excellent medium for the growth of pathogenic and non-pathogenic bacteria. This study aims to determine the characteristics of the bacterial genus in broiler chicken carcasses from supermarkets in Pontianak City. Based on the results of the study found 23 bacterial isolates in broiler chicken carcass samples from supermarkets in Pontianak City, which included members of the Aeromonas, Acetobacter, Alcaligenes, Amphibacillus, Bacillus, Brevibacterium, Camphylobacter, Carnobacterium, Erwinia,  Erysipelothrik, Eubacterium, Hafnia, Kluyvera, Klebsiella, Kurthia, Lactobacillus, Listeria, Proteus, Pseudomonas, Shigella, Sporolactobacillus, Serratia, and  Yersinia.

scholarly journals Antibiotic Susceptibility and Genotype Patterns of Escherichia coli, Klebsiella pneumonia and Pseudomonas aeruginosa Isolated from Urinary Tract Infected Patients

2010 ◽  
Vol 59 (3) ◽  
pp. 207-212 ◽  
Author(s):  
M.I. ABOU-DOBARA ◽  
M.A. DEYAB ◽  
E.M. ELSAWY ◽  
H.H. MOHAMED
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Urinary Tract ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Random Amplified Polymorphic Dna ◽  
Test Methods ◽  
Klebsiella Pneumonia ◽  
E Coli ◽  
Rapd Pcr

Thirty nine isolates of Escherichia coli, twenty two isolates of Klebsiella pneumoniae and sixteen isolates of Pseudomonas aeruginosa isolated from urinary tract infected patients were analyzed by antimicrobial susceptibility typing and random amplified polymorphic DNA (RAPD)-PCR. Antibiotic susceptibility testing was carried out by microdilution and E Test methods. From the antibiotic susceptibility, ten patterns were recorded (four for E. coli, three for K. pneumoniae and three for P. aeruginosa respectively). Furthermore, genotyping showed seventeen RAPD patterns (seven for E. coli, five for K. pneumoniae and five for P. aeruginosa respectively). In this study, differentiation of strains of E. coli, K. pneumoniae and P. aeruginosa from nosocomial infection was possible with the use of RAPD.

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