Comparison of three molecular typing methods in studies of transmission of Porphyromonas gingivalis

Steenbergen, T. J. M. van; Ménard, Christian; Tijhof, C. J.; Mouton, Christian; Graaff, J. de

doi:10.1099/00222615-39-6-416

Cited by 39 publications

(30 citation statements)

References 16 publications

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“…As the RAPD profiles have only a small number of bands and, consequently, are much easier to read than the corresponding REA patterns, they should be considered helpful for epidemiological and transmission studies of mutans and other oral streptococci, as already shown with P. gingivalis [27] and suggested for mutans streptococci [28,29].…”

Section: Discussionmentioning

confidence: 99%

“…They include DNA-DNA hybridisation [1,12,14], restriction endonuclease analysis (REA) [16][17][18][19], multilocus enzyme electrophoresis (MLEE) [20], ribotyping, PCR and colony hybridisation with DNA probes [21]. A PCR-based procedure of DNA fingerprinting, called arbitrarily primed PCR (AP-PCR) [22] or randomly amplified polymorphic DNA (RAPD) analysis [23], has been applied to strain identification and typing of oral bacteria [24][25][26][27], oral streptococci [28][29][30] and non-oral streptococci [31][32][33][34]. The present study used RAPD analysis for typing S. mutans and S. sobrinus strains and to explore the potential of this typing method to distinguish these from the other taxa that comprise the group of mutans streptococci.…”

Section: Introductionmentioning

confidence: 99%

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Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis

Truong¹,

Ménard

Mouton³

et al. 2000

Journal of Medical Microbiology

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The identification and classification of the non-haemolytic or viridans group of streptococci have long been recognised as difficult and unsatisfactory. Phenotypic and genotypic heterogeneity have resulted in ambiguous speciation, particularly with mutans streptococci and other oral streptococci. This study was done to determine whether random amplified polymorphic DNA (RAPD) analysis is useful to identify and even classify oral and other streptococci. DNA was prepared and purified from 25 strains of mutans streptococci including 11 reference strains of Streptococcus mutans, seven of S. sobrinus, three of S. rattus and one each of the four other species of the mutans group, together with 20 other reference species, mostly streptococci, and from 49 fresh isolates of mutans streptococci and of S. mutans from human saliva and dental plaque. DNA amplification was primed with each of three arbitrarily selected primers nine or 10 nucleotides in length. The amplified DNA fragments (amplicons) obtained were compared by agarose gel electrophoresis. Species-and strain-specific RAPD fingerprints were obtained not only from pure genomic DNA, but also from the supernates of crude cellular or colony extracts. Pending the analysis of numerous other strains, the data suggest that RAPD may be of value: (i) to distinguish the species S. mutans and S. sobrinus from each other and potentially from other species of oral streptococci, (ii) to differentiate and possibly classify oral streptococci and (iii) as a valuable tool in mutans streptococci epidemiology and transmission studies, by virtue of its rapidity, efficiency and reproducibility in generating genetic fingerprints of streptococcal isolates.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis

Truong¹,

Ménard

Mouton³

et al. 2000

Journal of Medical Microbiology

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“…These methods include restriction endonuclease analysis (10,27,31), restriction fragment length polymorphism (RFLP) (8), ribotyping (27,31), AP-PCR (1, 27, 31), multilocus enzyme electrophoresis (19), heteroduplex analysis (26), and fimA genotyping (23). The genotypic characterization of the P. gingivalis strains has revealed extensive heterogeneity.…”

Section: Discussionmentioning

confidence: 99%

“…In P. gingivalis, interspousal transmission was detected in 20 to 75% of cases by AP-PCR or a heteroduplex analysis of the ribosomal ISR. However, transmission from parent to child appeared to be uncommon (2,31). Nevertheless, the finding that concordance in the colonization of P. gingivalis was frequently observed for children and their mothers by detecting the presence of P. gingivalis by PCR assay may implicate the vertical transmission of this bacterium (30,33).…”

Section: Vol 42 2004mentioning

confidence: 99%

Use of Insertion Sequence Element IS 1126 in a Genotyping and Transmission Study of Porphyromonas gingivalis

et al. 2004

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Porphyromonas gingivalis is strongly associated with periodontal diseases and is regarded as one of the risk factors for periodontitis. Insertion sequence element IS1126-based PCR was used to investigate the genetic heterogeneity of P. gingivalis from periodontitis patients and to examine the frequency of the parent-child and spouse-spouse transmission. Two sets of IS1126-specific primers were used for the PCR. The inward primer set (PI1 and PI2), which amplifies the IS1126 fragment of approximately 690 bp, was used to identify P. gingivalis. The outward primer set (PI1RC and PI2RC), which is reverse complementary to PI1 and PI2, respectively, and amplifies the gene fragments between the adjacent IS1126 elements was used to characterize the genotypes of the P. gingivalis strains. PCR of P. gingivalis with PI1RC and PI2RC resulted in the production of two to seven amplicons, which showed a unique electrophoretic pattern in each strain (4 laboratory strains and 37 clinical isolates cultured from 12 patients with aggressive periodontitis). The usefulness of the method for transmission study was confirmed by detecting identical genotypes between the isolates and the plaque samples from which the isolates were cultured and between the plaque samples from different tooth sites in the same patient. Thirty probands with periodontal diseases and their thirty immediate family members were included in the transmission study. In 11 of 14 parent-child pairs (78.6%), P. gingivalis revealed an identical or similar band pattern, whereas 5 of 16 spouse pairs (31.25%) had this similarity. These results show that IS1126-based PCR for genotyping P. gingivalis has a highly discriminating potential with reproducible data and is a simple and reliable method for a transmission study.Porphyromonas gingivalis is a gram-negative, anaerobic, nonmotile short rod-shaped, black-pigmented bacterium. Studies on the epidemiology, the virulence factors, the host response, and the pathogenicity in animal models have provided strong evidence implicating this bacterium in the etiology of periodontitis (9, 10, 12). P. gingivalis is known to be a risk factor for periodontitis (3). In addition, there is accumulating evidence showing that an infection with P. gingivalis may predispose an individual to cardiovascular disease and cause the delivery of preterm infants (4,5,24). This emphasizes the importance of preventing an infection by this bacterium. In this respect, information on the transmission route of P. gingivalis between individuals is important for determining the risk factors for periodontitis and for developing strategies for preventing the transmission of the pathogen to uninfected individuals.Several studies on the transmission of periodontopathogens have been performed by detecting identical strains between related family members. The transmission of Actinobacillus actinomycetemcomitans and P. gingivalis has been reported by comparing either the serotypes (15, 32) or the genotypes. The transmission rate reported thus far varies from study ...

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“…P. endodontalis와 P. gingivalis는 근관감염의 급성증상과 밀접한 관련이 있었고, P. intermedia와 P. nigrescens는 증상과 관계없 이 관찰되었다 22,23) . 24,27,28) . 또한 PCR은 세균 의 대한 DNA sequence 자료가 부족하여 모든 세균에 대 해 적용할 수 없는 단점을 가지며 특정한 균에 대해서만 특 이적인 primer를 제작하여 동정하고자 하는 세균의 DNA 와 모든 의심되는 균의 primer를 반응시켜 증폭시키는 과 정이 필요하다.…”

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Detection of black-pigmented bacteria in infected root canals

Kwon

Kim

Kwak

et al. 2002

J Korean Acad Conserv Dent

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Black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections.The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P.nigrescens by using special potency disk test, filter paper spot test, 16S rRNA gene-directed PCR, and API 32A. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and/or apical periodontitis. Conventional laboratory methods were used for identification of the strains of black pigmented bacteria. Eighteen of 33 samples were positive for the growth of black-pigmented bacrteria. Five colonies were cultured from each pure cultured colonies from Brucella agar plate. Seventy seven colonies were positive for the growth of black-pigmented bacteria.Thirty three of 77(42.6%) were identifed as P. nigrescens, 10 of 77(12.9%)were P. gingivalis, 6 of 77(7.8%) were P. endodontalis, 10 of 77(12.9%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P. nigrescens was sensitive to kanamycin in special potency disk test.16S rRNA gene PCR and API test after rapid presumptative identification methods, such as special potency disk test and filter paper spot test, would be accurate detection methods for black-pigemented bac-Ⅰ. 서 론 치수 및 치근단 질환의 발병과 진행에 있어서 세균은 주요 한 원인인자로서 대부분의 근관감염은 혼합감염이고 특히 혐기성세균이 주를 이룬다 [1][2][3] . 이들 혐기성 세균중 blackpigmented bacteria는 치근단 질환의 주요 병인균으로 주 목받고 있으며, 악취, 동통, 누공 형성과 지속적인 임상증상 이 있는 경우나 만성 치근단 질환의 악화에 중요한 역할을 담당한다 [4][5][6][7][8] . 이전에 black-pigmented bacteria는. Shah와 Collins는 Bacteroides를 세 가지 속으로 나누어, saccharolytic nonpigmenting species로 구성된 Bacteroides, asaccharolytic black-pigmenting species로 구성된 Porphyromonas 종 및 saccharolytic black-pigmented species와 nonpigmenting species로 구성된 Prevotella로 구분하였다 9,10)

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Comparison of three molecular typing methods in studies of transmission of Porphyromonas gingivalis

Cited by 39 publications

References 16 publications

Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis

Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis

Use of Insertion Sequence Element IS 1126 in a Genotyping and Transmission Study of Porphyromonas gingivalis

Detection of black-pigmented bacteria in infected root canals

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