scholarly journals Genotypic Variations of Mutans Streptococci Isolated from Dental Caries by REP-PCR

2020 ◽  
Vol 17 (4) ◽  
pp. 1133
Author(s):  
Hamzah Abdulrahman Salman ◽  
R. Senthilkumar
Keyword(s):  
Pcr Amplification ◽  
Oral Bacteria ◽  
Mutans Streptococci ◽  
Streptococcus Sobrinus ◽  
Clinical Strains ◽  
Core Species ◽  
Banding Profile ◽  
Common Band ◽  
Species Specific ◽  
Reference Strains

Mutans streptococci (MS) are a group of oral bacteria considered as the main cariogenic organisms. MS consists of several species of genus Streptococcus which are sharing similar phenotypes and genotypes. The aim of this study is to determine the genetic diversity of the core species of clinical strains of Streptococcus mutans, Streptococcus sobrinus and Streptococcus downei by using repitative extragenic palindromic (REP) primer. The DNA of the clinical strains of S. mutans (n=10), S. sobrinus (n=05) and S. downei (n=04) have been employed in the present study, which have been previously isolated from caries active subjects. The DNA of the clinical and reference strains was subjected to PCR amplification using REP primer. The phylogenetic dendrogram is constructed from the REP PCR banding profile by neighbour-joining method using PyElph 1.4 software. The size of the DNA amplicons generated by using REP primer were S. mutans (1500 bp to 250 bp), S. sobrinus (6000 bp to 250 bp) and S. downei (5000 bp to 400 bp). The results present common band at 480 bp in all the clinical strains of S. sobrinus. The current study is the first to demonstrate the genetic variety of S. sobrinus and S. downei by using REP primer. REP-PCR have been found to be a powerful method to study the molecular diversity of S. mutans, S. sobrinus and S. downei. Additionally, further studies are suggested to analyze the species specific bands and also to find the possibility to produce a new specific primer for S. sobrinus.

The Use of High - Sucrose Culture Media for the Identification of Oral Streptococci in Infant- Mother Pairs

2017 ◽  
Vol 68 (11) ◽  
pp. 2691-2693
Author(s):  
Krisztina Martha ◽  
Cristina Bica ◽  
Edva Anna Frunda
Keyword(s):  
Culture Media ◽  
Bacterial Flora ◽  
Mutans Streptococci ◽  
Resistant Bacteria ◽  
Oral Streptococci ◽  
Streptococcus Sobrinus ◽  
Sugar Intake ◽  
Tooth Surfaces ◽  
Refined Carbohydrates ◽  
High Sucrose

By the end of the 60�s, the theory that refined carbohydrates promotes the absorption of saccharolytic Gram-positive microbial species on the tooth surfaces has become generally. Mutans streptococci (Streptococcus mutans and Streptococcus sobrinus) were key players in this theory. On agar plates, Str. mutans produces small, circular colonies, in the presence of glucose, and in the presence of sucrose large, sticky, gelatinous colonies. This gelatinous texture is due to the shell material: mutant 1 � 3 glucose polymers and dextran 1 �! 6 glucose polymers. Str. mutans are able to survive in the oral cavity with a pH lower than 5.5. That is why consecutive multiple sugar intake promotes the colonization of Str. mutans, which results in dental caries in stagnant zones. As oral pH is continuously shifted to acid, more acid-resistant bacteria appear. Our aim was to identify species in infant-mother pair gingival crevicular bacterial flora, which can be detected on high-sucrose culture media and to underline the jeopardy of vertical oral contamination from mother to infant.

ISSR-Derived Species-Specific SCAR Marker for Rapid and Accurate Authentication of Ocimum tenuiflorum L.

Planta Medica ◽  
2017 ◽  
Vol 84 (02) ◽  
pp. 117-122 ◽  
Author(s):  
Amit Kumar ◽  
Vereena Rodrigues ◽  
Priyanka Mishra ◽  
Kuppusamy Baskaran ◽  
Ashutosh Shukla ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
High Volume ◽  
Inter Simple Sequence Repeat ◽  
Rapid Identification ◽  
Medicinal Species ◽  
Accurate Identification ◽  
Sequence Characterized Amplified Region ◽  
Medicinal Value ◽  
Ocimum Tenuiflorum ◽  
Species Specific

Abstract Ocimum tenuiflorum has been widely used in traditional medicine and has high medicinal value. High volume trade of this potential medicinal plant species led to unscrupulous adulteration of both crude drugs as well as formulations. Morphology-based authentication is difficult in cases of incomplete or damaged samples and in dried herbal materials. In such cases, PCR-based molecular methods may aid in accurate identification. The present study aimed at developing species-specific DNA marker(s) for the authentication of O. tenuiflorum. A species-specific amplicon (279 bp) generated through an inter-simple sequence repeat marker (UBC 835) in all individuals of O. tenuiflorum was cloned, sequenced, and a primer pair was developed (designated as CIM-OT-835F/CIM-OT-835R). The newly developed sequence characterized amplified region marker was validated through PCR amplification in all available seven species of Ocimum, and its specificity for O. tenuiflorum was confirmed with the consistent generation of an amplicon of 177 bp. The developed marker can be used for accurate and rapid identification of the species for certification purposes and will be useful in quality control of medicinal preparations containing this important medicinal species.

scholarly journals Adherence of mutans streptococci to other oral bacteria.

1990 ◽  
Vol 58 (6) ◽  
pp. 1738-1743 ◽  
Author(s):  
R J Lamont ◽  
B Rosan
Keyword(s):  
Oral Bacteria ◽  
Mutans Streptococci

scholarly journals Molecular Characterization of Yarrowia lipolytica and Candida zeylanoides Isolated from Poultry

2000 ◽  
Vol 66 (10) ◽  
pp. 4340-4344 ◽  
Author(s):  
T. Deak ◽  
J. Chen ◽  
L. R. Beuchat
Keyword(s):  
Yarrowia Lipolytica ◽  
Rapd Analysis ◽  
Restriction Analysis ◽  
Pcr Amplification ◽  
Yeast Cells ◽  
Chromosomal Dna ◽  
Intergenic Sequences ◽  
Species Specific ◽  
Candida Zeylanoides ◽  
Its Pcr

ABSTRACT Yeast isolates from raw and processed poultry products were characterized using PCR amplification of the internally transcribed spacer (ITS) 5.8S ribosomal DNA region (ITS-PCR), restriction analysis of amplified products, randomly amplified polymorphic DNA (RAPD) analysis, and pulsed-field gel electrophoresis (PFGE). ITS-PCR resulted in single fragments of 350 and 650 bp, respectively, from eight strains of Yarrowia lipolytica and seven strains of Candida zeylanoides. Digestion of amplicons with HinfI andHaeIII produced two fragments of 200 and 150 bp fromY. lipolytica and three fragments of 350, 150, and 100 bp from C. zeylanoides, respectively. Although these fragments showed species-specific patterns and confirmed species identification, characterization did not enable intraspecies typing. Contour-clamped heterogeneous electric field PFGE separated chromosomal DNA of Y. lipolytica into three to five bands, most larger than 2 Mbp, whereas six to eight bands in the range of 750 to 2,200 bp were obtained from C. zeylanoides. Karyotypes of both yeasts showed different polymorphic patterns among strains. RAPD analysis, using enterobacterial repetitive intergenic sequences as primers, discriminated between strains within the same species. Cluster analysis of patterns formed groups that correlated with the source of isolation. For ITS-PCR, extraction of DNA by boiling yeast cells was successfully used.

scholarly journals Effects of Caries Activity on Compositions of Mutans Streptococci in Saliva-Induced Biofilm Formed on Bracket Materials

Materials ◽  
2020 ◽  
Vol 13 (21) ◽  
pp. 4764
Author(s):  
Bum-Soon Lim ◽  
Bo-Hyun Kim ◽  
Won-Jun Shon ◽  
Sug-Joon Ahn
Keyword(s):  
Polymerase Chain Reaction ◽  
Streptococcus Mutans ◽  
Mutans Streptococci ◽  
Growth Media ◽  
Streptococcus Sobrinus ◽  
Chain Reaction ◽  
Caries Activity ◽  
Material Type ◽  
Polymerase Chain ◽  
Total Bacteria

This study aimed to investigate effects of caries activity on composition of mutans streptococci in saliva-induced biofilms formed on bracket materials. Three bracket materials were used as specimens: ceramic, metal, and plastic. After saliva was collected using a spitting method from caries-active (CA, decayed, missing, filled teeth (DMFT) score ≥ 10) and caries-free (CF, DMFT score = 0) subjects, saliva was mixed with growth media in a proportion of 1:10. The saliva solution was then incubated with each bracket material. After a saliva-induced biofilm was developed on the surface of the bracket material, the amounts of total bacteria and mutans streptococci were determined using real-time polymerase chain reaction. The results showed that biofilms from CA saliva contained more mutans streptococci but less total bacteria than biofilms from CF saliva, regardless of material type. Adhesion of total bacteria to ceramic was higher than to plastic, regardless of caries activity. Mutans streptococci adhered more to ceramic than to metal and plastic in both biofilms from CA and CF saliva, but there was a difference in adhesion between Streptococcus mutans and Streptococcus sobrinus. The amount of S. mutans was higher than that of S. sobrinus in biofilms from CA saliva despite similar amounts of the two strains in biofilms from CF saliva. The stronger adhesion of S. mutans to ceramic than to metal and plastic was more evident in biofilms from CA saliva than in biofilms from CF saliva. This study suggests that caries activity and material type significantly influenced composition of mutans streptococci in biofilms formed on bracket materials.

scholarly journals New Pyrimidinone-Fused 1,4-Naphthoquinone Derivatives Inhibit the Growth of Drug Resistant Oral Bacteria

Biomedicines ◽  
2020 ◽  
Vol 8 (6) ◽  
pp. 160
Author(s):  
Kyungmin Kim ◽  
Daseul Kim ◽  
Hyunjin Lee ◽  
Tae Hoon Lee ◽  
Ki-Young Kim ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Physiological Activity ◽  
Phenyl Group ◽  
Fusobacterium Nucleatum ◽  
Antimicrobial Activities ◽  
Oral Bacteria ◽  
Drug Resistant ◽  
Streptococcus Sobrinus ◽  
Disk Diffusion Assay ◽  
Drug Resistant Strains

Background: Dental caries is considered to be a preventable disease, and various antimicrobial agents have been developed for the prevention of dental disease. However, many bacteria show resistance to existing agents. Methods/Principal Findings: In this study, four known 1,4-naphthoquinones and newly synthesized 10 pyrimidinone-fused 1,4-naphthoquinones, i.e. KHQ 701, 702, 711, 712, 713, 714, 715, 716, 717 and 718, were evaluated for antimicrobial activity against Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Actinomyces viscosus and Fusobacterium nucleatum. Pyrimidinone-fused 1,4-naphthoquinones were synthesized in good yields through a series of chemical reactions from a commercially available 1,4-dihydroxynaphthoic acid. MIC values of KHQ 711, 712, 713, 714, 715, 716, 717 and 718 were 6.25–50 μg/mL against E. faecalis (CCARM 5511), 6.25–25 μg/mL against E. faecium (KACC11954) and S. aureus (CCARM 3506), 1.56–25 μg/mL against S. epidermidis (KACC 13234), 3.125–100 μg/mL against S. mutans (KACC16833), 1.56–100 μg/mL against S. sobrinus (KCTC5809) and P. gingivalis (KCTC 5352), 3.125–50 μg/mL against A. viscosus (KCTC 9146) and 3.125–12.5 μg/mL against F. nucleatum (KCTC 2640) with a broth microdilution assay. A disk diffusion assay with KHQ derivatives also exhibited strong susceptibility with inhibition zones of 0.96 to 1.2 cm in size against P. gingivalis. Among the 10 compounds evaluated, KHQ 711, 712, 713, 715, 716 and 717 demonstrated strong antimicrobial activities against the 9 types of pathogenic oral bacteria. A pyrimidin-4-one moiety comprising a phenyl group at the C2 position and a benzyl group at the N3 position appears to be essential for physiological activity. Conclusion/Significance: Pyrimidinone-fused 1,4-naphthoquinones synthesized from simple starting compounds and four known 1,4-naphthoquinones were synthesized and showed strong antibacterial activity to the 9 common oral bacteria. These results suggest that these derivatives should be prospective for the treatment of dental diseases caused by oral bacteria, including drug-resistant strains.

scholarly journals Species-Specific and Ubiquitous-DNA-Based Assays for Rapid Identification of Staphylococcus aureus

1998 ◽  
Vol 36 (3) ◽  
pp. 618-623 ◽  
Author(s):  
Francis Martineau ◽  
François J. Picard ◽  
Paul H. Roy ◽  
Marc Ouellette ◽  
Michel G. Bergeron
Keyword(s):  
Staphylococcus Aureus ◽  
Genomic Library ◽  
Antibiotic Resistance Genes ◽  
Pcr Amplification ◽  
Rapid Identification ◽  
Clinical Microbiology Laboratory ◽  
Chromosomal Dna ◽  
Pcr Assay ◽  
Serious Infections ◽  
Species Specific

Staphylococcus aureus is the cause of serious infections in humans, including endocarditis, deep-seated abscesses, and bacteremia, which lead to toxic and septic shock syndromes. Rapid and direct identification of this bacterium specifically and ubiquitously directly from clinical specimens would be useful in improving the diagnosis of S. aureus infections in the clinical microbiology laboratory. A wide variety of kits based on biochemical characteristics efficiently identify S. aureus, but the rapidity and the accuracy of each of these methods combined with testing of clinically relevant antibiotic resistance genes need to be improved. On the basis of hybridization assays with randomly selected clones from an S. aureus genomic library, we have identified a chromosomal DNA fragment which is specific for S. aureus and which detected all 82 S. aureus isolates tested. This 442-bp fragment was sequenced and was used to design a set of PCR amplification primers. The PCR assay was also specific and ubiquitous for the identification from bacterial cultures of 195 clinical strains of S. aureus isolated from a variety of anatomical sites and obtained from hospitals throughout the world. The PCR assay that we have developed is simple and can be performed in about 1 h. This DNA-based test provides a novel diagnostic tool for the diagnosis of S. aureus infections.

PCR-based molecular discrimination betweenMaritrema eroliaeandProbolocoryphe uca(Digenea: Microphallidae) in Kuwait Bay

2013 ◽  
Vol 88 (2) ◽  
pp. 177-182
Author(s):  
W.Y. Al-Kandari ◽  
S.A. Al-Bustan ◽  
M. Alnaqeeb ◽  
A.M. Isaac
Keyword(s):  
Fragment Length Polymorphism ◽  
Pcr Amplification ◽  
Direct Pcr ◽  
Specific Primers ◽  
Kuwait Bay ◽  
Future Studies ◽  
Larval Stages ◽  
Marine Snails ◽  
Pcr Rflp ◽  
Species Specific

AbstractMicrophallid trematodes are common parasites in marine snails and crustacean hosts at Kuwait Bay. The larval stages of two microphallids,Maritrema eroliaeandProbolocoryphe uca, are difficult to differentiate morphologically. In this study, two PCR-based techniques were established for quick and accurate discrimination between the larval stages of the two microphallid species, employing restriction fragment length polymorphism (PCR-RFLP) and species-specific primers. Both techniques utilized nucleotide differences in the second internal transcribed region (ITS2) of the ribosomal DNA (rDNA) in the two species. For the PCR-RFLP technique, restriction enzymeAvaII was selected and it generated different restriction profiles among the two microphallids. In addition, species-specific primers were prepared for each microphallid species that amplified distinctive fragments. Both techniques showed that the larval stages of the two microphallid species can be identified accurately. However, direct PCR amplification using species-specific primers was more advantageous than the PCR-RFLP technique since it allowed rapid and specific discrimination between the two species. This technique provides a useful tool that can be used in future studies for the study of the distribution of microphallid species and their definitive hosts at different localities of Kuwait Bay.

scholarly journals The condundrum of feline trichomonosis – The more we learn the ‘trickier’ it gets

2017 ◽  
Vol 19 (3) ◽  
pp. 261-274 ◽  
Author(s):  
Jody L Gookin ◽  
Katherine Hanrahan ◽  
Michael G Levy
Keyword(s):  
Chronic Diarrhea ◽  
Safety Margin ◽  
Pcr Amplification ◽  
Evidence Base ◽  
Domestic Cat ◽  
Tritrichomonas Foetus ◽  
Specific Primers ◽  
Clinical Resistance ◽  
Comprehensive Examination ◽  
Species Specific

Practical relevance: Trichomonosis of the large intestine of the cat was described as a cause of chronic diarrhea over 20 years ago. The trichomonad was identified as Tritrichomonas foetus, with a genotype that is distinct from venereal T foetus of cattle. Clinical challenges: Despite multiple means for diagnosis of the infection, including light microscopy, protozoal culture and PCR amplification using species-specific primers, tests with even greater sensitivity are needed. Feline trichomonosis is resistant to all commonly used antiprotozoal drugs. Ronidazole is currently the only drug demonstrated to be effective in eliminating the infection from cats; however, this drug has a narrow safety margin and clinical resistance is increasingly recognized. The more we learn about trichomonosis in cats, the more complicated and controversial the infection has become, ranging from what we should call the organism to whether we should even bother trying to treat it. Global importance: Feline trichomonosis is recognized to occur worldwide and is regarded as one of the most common infectious causes of colitis in the domestic cat. The infection is widespread in catteries and shelters; and, while remission of diarrhea may occur over time, persistence of the infection is common. Evidence base: This review provides a comprehensive examination of what is currently known about feline trichomonosis and pinpoints areas, based on the authors’ opinion, where further research is needed.

Sign in / Sign up

Export Citation Format

Share Document