牙周炎患者唾液中伴放线放线杆菌的检出状况分析

目的 检测不同类型牙周炎患者唾液中的伴放线放线杆菌(Actinobacillusactinomycetemcomitans,Aa),探讨唾液和集合龈下菌斑中Aa检出率的差异以及唾液中Aa的存在状况与牙周临床指标的关系. 方法 收集50例侵袭性牙周炎(aggressive periodontitis,AgP)患者、48例慢性牙周炎(chronic periedontitis,CP)患者和25例非牙周炎者的非刺激性全唾液和集合龈下菌斑,应用聚合酶链反应(PcR)技术检测两种样本中的Aa. 结果 Aa在AgP患者唾液中的检出率(32%)显著高于非牙周炎者(4%)和CP患者(15%),差异均有统计学意义(P<0.01,P<0.05),同时Aa在AgP患者唾液中的检出率也显著高于集合龈下菌斑样本(16%),差异亦有统计学意义(P<0.05).年龄≤30岁是唾液中存在Aa的危险指征(OR=3.23,P<0.05);出血指数≥3的位点超过70%与唾液中存在Aa有关(OR=19.21,P<0.01). 结论 AgP患者唾液样本中Aa的检出率明显高于集合龈下菌斑样本,亦高于CP患者和非牙周炎者,提示Aa可能参与AgP的发生和发展.     

实时荧光定量PCR检测牙周炎患者龈下菌斑的分布

目的 应用实时荧光定量PCR技术探索侵袭性牙周炎（aggressive periodontitis,AgP）、慢性牙周炎（chronic periodontitis,CP）患者龈下菌斑中伴放线聚集杆菌(A. actinomycetemcomitans,Aa)、牙龈卟啉单胞菌（P. gingivalis,Pg）的分布规律。方法 采集32例AgP、33例CP、32例牙周健康者的龈下菌斑,构建含有2种待测细菌基因片段的重组质粒,建立定量标准,采用TaqManMGB探针实时荧光定量PCR方法检测样本中细菌数量。结果 本实验构建的引物及TaqManMGB探针特异性及敏感性较好。AgP组龈下菌斑Aa的检出率高于CP组（P＜0.01）,但2种细菌数量在组间无显著差异,两组内Pg的检出率及数量都明显高于Aa（P＜0.001）,另外AgP组Aa的数量、CP组Pg数量与牙周探诊深度密切相关（P＜0.01及P＜0.001）。结论 龈下菌斑Aa的检出率可能与牙周炎类型存在一定关联,Aa可能并不是中国人群样本AgP患者龈下菌斑的优势菌,实时荧光定量PCR对牙周病学研究有广泛应用前景。     

伴放线放线杆菌在龈下菌斑和颊黏膜分布研究

目的:比较伴放线放线杆菌(actinobac illus actinomycetem com itans,A.a)在不同类型牙周炎患者龈下菌斑和颊黏膜中的分布。方法:通过聚合酶链反应(polym erase chain reaction,PCR)对侵袭性牙周炎患者(AgP)、慢性牙周炎患者(CP)、牙周健康者口腔龈下菌斑和颊黏膜中的A.a进行检测,分析该菌分别在两部位的相对含量。结果:AgP组菌斑和颊黏膜样本中A.a阳性检出率均为41.7%,分别高于CP组(菌斑16.7%、颊黏膜10.0%)和牙周健康组(菌斑和颊黏膜均为0%)。AgP组A.a在菌斑和颊黏膜的相对含量分别为38.5%和22.2%,高于CP组(菌斑19%、颊黏膜12.75%)。结论:A.a不仅存在于龈下菌斑中,也能够粘附于颊黏膜;A.a是AgP的主要优势菌也参与了CP的菌群组成。     

侵袭性牙周炎与慢性牙周炎病变位点中人巨细胞病毒的定量检测

目的 应用实时荧光定量PCR方法 检测侵袭性牙周炎(AgP)及慢性牙周炎(CP)患者龈下样本中人巨细胞病毒(HCMV)的DNA载量,探讨HCMV感染与牙周炎之间的关系.方法 选择18例AgP患者、24例CP患者及15例牙周健康对照者,收集龈下样本114例.构建含有HCMV高保守片段的重组质粒,制备标准品DNA模板,建立...     

2型糖尿病患者慢性牙周炎龈下菌群分析

目的 :比较 2型糖尿病 (DM )慢性牙周炎与非糖尿病慢性牙周炎的龈下菌群 ,为牙周炎的防治提供参考。方法 :厌氧菌培养、PCR检测技术。结果 :①糖尿病慢性牙周炎与非糖尿病慢性牙周炎患者临床指标无明显差别 (P >0 .0 5 )。②糖尿病慢性牙周炎的龈下菌群与非糖尿病慢性牙周炎相似 ,以厌氧菌为主 ,兼有少量口腔链球菌 ;产黑色素类杆菌、核梭杆菌、二氧化碳噬纤维菌、中间普氏菌为牙周优势菌 (P <0 .0 1)。③糖尿病慢性牙周炎的龈下菌群厌氧菌总数、口腔链球菌总数、口腔乳杆菌及核梭杆菌数量少于非糖尿病慢性牙周炎者。结论 :2型糖尿病慢性牙周炎与非糖尿病慢性牙周炎龈下菌群相似 ,致病菌数量低于非糖尿病牙周炎患者。     

侵袭性牙周炎病原微生物的检测

目的检测侵袭性牙周炎（AgP）患者和牙周健康者龈下菌斑中的7种病原微生物,旨在寻找AgP的主要致病微生物.方法应用以16S rRNA为基础的聚合酶链反应（PCR）技术,检测55例AgP患者和17名健康对照者龈下菌斑中的7种牙周病原微生物：伴放线放线杆菌（Aa）,牙龈卟啉单胞菌（Pg）,福赛坦氏菌（Tf）,牙密螺旋体（Td）,直肠弯曲杆菌（Cr）,中间普氏菌（Pi）,变黑普氏菌（Pn）.结果55例AgP患者中仅有1例检测出Aa,而在健康对照者中未检出该菌.Pg、Tf、Td和Cr在AgP组的检出率分别为81.8%、83.6%、80.0%和81.8%,显著高于健康对照者（17.6%、11.8%、5.9%、29.4%）,差异有统计学意义（P＜0.01）.结论Pd、Tf、Td和Cr 4种微生物在AgP患者中有较高的检出率,提示它们的共同定植可能在AgP中起重要作用.     

伴2型糖尿病的慢性牙周炎牙周可疑致病菌的检测

目的 检测伴2型糖尿病(diabetes mellitus,DM)的慢性牙周炎(chronic periodontitis,CP)患者龈下菌斑中牙周可疑致病菌的种类和构成,从微生物学角度探讨牙周炎与DM的相互作用机制.方法 采集伴2型DM的CP患者154例(DM组)、不伴DM的单纯CP患者120例(CP组)及40名全身及牙周健康者(N组)的龈下集合菌斑,传统酚-氯仿法提取菌斑DNA,以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg),伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa),具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf),齿垢密螺旋体(Treponema denticola,Td)为目标菌,应用以16SrRNA为基础的聚合酶链反应(PCR)技术对龈下菌群进行检测.结果 Pg、Aa、Fn、Pi、Tf、Td在DM组中均可检出;与CP组相比,在性别、年龄、牙周状况基本一致的情况下,轻度牙周炎者DM组Pi的检出率为35%(8/23),CP组为65%(13/20),两组间差异有统计学意义(P<0.05);重度牙周炎者DM组Pg、Aa、Tf的检出率分别为78%(72/92)、27%(25/92)、67%(62/92),CP组分别为58%(35/60)、17%(10/60)、43%(26/60),DM组均显著高于CP组,差异有统计学意义(P<0.05).同时,DM组Aa、Tf PCR产物的平均灰度值(average gradation,AVG)比值显著高于CP组,Pi的AVG比值明显低于CP组,P<0.05.结论 与单纯CP相比,伴2型DM的牙周炎患者龈下菌斑中Pg、Aa、Tf的数量增多,Pi的数量减少.     

伴2型糖尿病的慢性牙周炎牙周可疑致病菌的检测

目的 检测伴2型糖尿病(diabetes mellitus,DM)的慢性牙周炎(chronic periodontitis,CP)患者龈下菌斑中牙周可疑致病菌的种类和构成,从微生物学角度探讨牙周炎与DM的相互作用机制.方法 采集伴2型DM的CP患者154例(DM组)、不伴DM的单纯CP患者120例(CP组)及40名全身及牙周健康者(N组)的龈下集合菌斑,传统酚-氯仿法提取菌斑DNA,以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg),伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa),具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf),齿垢密螺旋体(Treponema denticola,Td)为目标菌,应用以16SrRNA为基础的聚合酶链反应(PCR)技术对龈下菌群进行检测.结果 Pg、Aa、Fn、Pi、Tf、Td在DM组中均可检出;与CP组相比,在性别、年龄、牙周状况基本一致的情况下,轻度牙周炎者DM组Pi的检出率为35%(8/23),CP组为65%(13/20),两组间差异有统计学意义(P<0.05);重度牙周炎者DM组Pg、Aa、Tf的检出率分别为78%(72/92)、27%(25/92)、67%(62/92),CP组分别为58%(35/60)、17%(10/60)、43%(26/60),DM组均显著高于CP组,差异有统计学意义(P<0.05).同时,DM组Aa、Tf PCR产物的平均灰度值(average gradation,AVG)比值显著高于CP组,Pi的AVG比值明显低于CP组,P<0.05.结论 与单纯CP相比,伴2型DM的牙周炎患者龈下菌斑中Pg、Aa、Tf的数量增多,Pi的数量减少.     

伴2型糖尿病的慢性牙周炎牙周可疑致病菌的检测

目的 检测伴2型糖尿病(diabetes mellitus,DM)的慢性牙周炎(chronic periodontitis,CP)患者龈下菌斑中牙周可疑致病菌的种类和构成,从微生物学角度探讨牙周炎与DM的相互作用机制.方法 采集伴2型DM的CP患者154例(DM组)、不伴DM的单纯CP患者120例(CP组)及40名全身及牙周健康者(N组)的龈下集合菌斑,传统酚-氯仿法提取菌斑DNA,以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg),伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa),具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf),齿垢密螺旋体(Treponema denticola,Td)为目标菌,应用以16SrRNA为基础的聚合酶链反应(PCR)技术对龈下菌群进行检测.结果 Pg、Aa、Fn、Pi、Tf、Td在DM组中均可检出;与CP组相比,在性别、年龄、牙周状况基本一致的情况下,轻度牙周炎者DM组Pi的检出率为35%(8/23),CP组为65%(13/20),两组间差异有统计学意义(P<0.05);重度牙周炎者DM组Pg、Aa、Tf的检出率分别为78%(72/92)、27%(25/92)、67%(62/92),CP组分别为58%(35/60)、17%(10/60)、43%(26/60),DM组均显著高于CP组,差异有统计学意义(P<0.05).同时,DM组Aa、Tf PCR产物的平均灰度值(average gradation,AVG)比值显著高于CP组,Pi的AVG比值明显低于CP组,P<0.05.结论 与单纯CP相比,伴2型DM的牙周炎患者龈下菌斑中Pg、Aa、Tf的数量增多,Pi的数量减少.     

慢性牙周炎合并糖尿病患者唾液及龈下菌斑中牙周致病菌检测临床研究

目的    探讨慢性牙周炎（CP）合并2型糖尿病（T2DM）患者与单纯CP患者唾液及龈下菌斑中牙周致病菌的差异性。方法    选择河南大学第一附属医院口腔科2012年8月至2015年9月伴T2DM的CP患者40例为CP+T2DM组,同时选取背景相似的慢性CP患者65例为CP组,并选取健康体检者35名作为对照组,采用聚合酶链反应（PCR）检测各组受试者唾液及龈下菌斑中牙龈卟啉单胞菌（Pg）、放线共生放线杆菌（Aa）、具核梭杆菌（Fn）、福塞坦菌（Tf）、中间型普里沃菌（Pi）、齿垢密螺旋体（Td）等可疑致病菌种类和相对含量。结果    3组受试者唾液及龈下菌斑中Pg、Aa和Tf检出率比较差异有统计学意义（P＜0.05）,而Fn、Pi和Td检出率差异无统计学意义（P＞0.05）,3组受试者唾液及龈下菌斑中Fn检出率均最高,CP+T2DM组唾液及龈下菌斑中Pg、Aa、Tf检出率均显著高于对照组和CP组（P＜0.05）,而CP组仅Pg检出率显著高于对照组（P＜0.05）,CP+T2DM组和CP组唾液及龈下菌斑中Pg、Aa和Tf相对含量均显著高于对照组（P＜0.05）,且CP+T2DM组唾液及龈下菌斑中Aa和Tf相对含量显著高于CP组,差异有统计学意义（P＜0.05）。结论    CP患者和伴糖尿病的CP患者牙周可疑致病菌的种类和数量存在明显差异,CP+T2DM组患者Pg、Aa和Tf检出率更高,可能与糖尿病关系更为密切。     

Immunoglobulin G subclass antibody profiles in Porphyromonas gingivalis-associated aggressive and chronic periodontitis patients

BACKGROUND/AIMS: The immunoglobulin G (IgG) antibody response is considered to be protective and beneficial for the control of periodontal lesions. This study analysed IgG subclass antibody levels of Porphyromonas gingivalis in patients with both aggressive periodontitis (AgP) and chronic periodontitis (CP). METHODS: Subgingival plaque and peripheral blood samples were collected from patients with localized AgP (n = 13), generalized AgP (n = 28) and generalized CP (n = 27) and from 14 periodontally healthy controls. P. gingivalis was identified in subgingival pockets using a polymerase chain reaction. Simultaneously, serum IgG subclass antibody against P. gingivalis whole cells/P. gingivalis fimbriae were measured using enzyme-linked immunosorbent assay. RESULTS: P. gingivalis was frequently detected in periodontitis patients. Anti-P. gingivalis whole cell IgG1 was elevated in all P. gingivalis-positive patients in the three periodontitis groups. Although increased anti-P. gingivalis IgG1 was also observed in the bacterium-positive healthy controls, the level was lower than that found in the three periodontitis groups. Levels of IgG1, IgG2, IgG3 and IgG4 to P. gingivalis did not differ among bacterium-positive patients in the three periodontitis groups; a significant increase of IgG2 level was not observed in localized AgP. Anti-fimbriae IgG subclass levels of IgG1, IgG2 and IgG4 did not differ among bacterium-positive subjects in all groups, while the anti-fimbriae IgG3 level in generalized CP was significantly higher than that in localized and generalized AgP. CONCLUSIONS: P. gingivalis infection elicited an IgG subclass antibody response in both periodontitis patients and healthy subjects, while higher anti-P. gingivalis IgG1 levels were found in the three periodontitis groups compared with the healthy control group.     

Immunoglobulin G subclass antibody profiles in Porphyromonas gingivalis‐associated aggressive and chronic periodontitis patients

Background/aims: The immunoglobulin G (IgG) antibody response is considered to be protective and beneficial for the control of periodontal lesions. This study analysed IgG subclass antibody levels of Porphyromonas gingivalis in patients with both aggressive periodontitis (AgP) and chronic periodontitis (CP). Methods: Subgingival plaque and peripheral blood samples were collected from patients with localized AgP (n = 13), generalized AgP (n = 28) and generalized CP (n = 27) and from 14 periodontally healthy controls. P. gingivalis was identified in subgingival pockets using a polymerase chain reaction. Simultaneously, serum IgG subclass antibody against P. gingivalis whole cells/P. gingivalis fimbriae were measured using enzyme‐linked immunosorbent assay. Results: P. gingivalis was frequently detected in periodontitis patients. Anti‐P. gingivalis whole cell IgG1 was elevated in all P. gingivalis‐positive patients in the three periodontitis groups. Although increased anti‐P. gingivalis IgG1 was also observed in the bacterium‐positive healthy controls, the level was lower than that found in the three periodontitis groups. Levels of IgG1, IgG2, IgG3 and IgG4 to P. gingivalis did not differ among bacterium‐positive patients in the three periodontitis groups; a significant increase of IgG2 level was not observed in localized AgP. Anti‐fimbriae IgG subclass levels of IgG1, IgG2 and IgG4 did not differ among bacterium‐positive subjects in all groups, while the anti‐fimbriae IgG3 level in generalized CP was significantly higher than that in localized and generalized AgP. Conclusions: P. gingivalis infection elicited an IgG subclass antibody response in both periodontitis patients and healthy subjects, while higher anti‐P. gingivalis IgG1 levels were found in the three periodontitis groups compared with the healthy control group.     

冠心病患者牙周状况及牙周细菌的检测

目的探讨冠心病患者牙周状况与牙龈卟啉单胞菌(Pg)和伴放线放线杆菌(Aa)的关系。方法试验分4组:冠心病伴慢性牙周炎组(A组)34例,冠心病组(B组)28例,慢性牙周炎组(C组)30例和对照组(D组)31例。记录各组牙周状况,取龈下菌斑进行Pg和Aa的分离培养及生化鉴定,检测牙周袋或龈沟内Pg和Aa阳性率。结果牙周状况A组比B组、C组差,C组比D组差。Pg、Aa阳性率A组比B组高,C组比D组高。结论冠心病患者的牙周健康状况较非冠心病患者差,提示慢性牙周炎可能是冠心病的又一危险因素。     

Predominant microflora of severe, moderate and minimal periodontal lesions in young adults with rapidly progressive periodontitis

The purpose of this investigation was to study the microflora of severe, moderate and minimal periodontal lesions, in young adults with rapidly progressive periodontitis (RPP). Subgingival plaque samples were taken from 142 periodontal lesions in 10 young adults aging 25 to 35 years. The examination of the subgingival microflora indicated that certain species, including Porphyromonas gingivalis, Bacteroides forsythus, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans , and Campylobacter species were found to be predominant in severe periodontal lesions. B. forsythus, P. gingivalis, Prevotella intermedia, F. nucleatum, Capnocytophaga ochracea , were predominant in medium lesions while Streptococcus species and Actinomyces species, C. ochracea, Haemophilus segnis and Veillonella parvula , were found in higher levels in minimal periodontal lesions.     

Prevalence of periodontopathic bacteria in aggressive periodontitis patients in a Chilean population

BACKGROUND: Actinobacillus actinomycetemcomitans is considered a major etiologic agent of aggressive periodontitis (AgP). Other periodontopathic bacteria such as Porphyromonas gingivalis are also suspected of participating in aggressive periodontitis although the evidence to support this is controversial. The aim of the present study was to determine the prevalence of eight periodontopathic bacteria in Chilean patients with AgP. METHODS: Subgingival plaque samples were collected from 36 aggressive, 30 localized, and six generalized periodontitis patients. Samples from 17 advanced chronic periodontitis (CP) patients were taken as controls. Samples collected from the four deepest periodontal pockets in each patient were pooled in prereduced transport fluid (RTF) and cultured. Periodontal bacteria were primarily identified by colony morphology under stereoscopic microscope and rapid biochemical tests. The identity of some bacterial isolates was confirmed by colony polymerase chain reaction (PCR). RESULTS: AgP showed a significatively higher prevalence of C. rectus than CP (P = 0.036). The only statistical difference found was for C. rectus. Patients with AgP showed a higher, but not statistically significant, prevalence of P. gingivalis, E. corrodens, P. micros, and Capnocytophaga sp. A similar prevalence in both groups of patients was observed for F. nucleatum and P. intermedia/nigrescens, and A. actinomycetemcomitans was less prevalent in AgP than CP patients. In localized AgP, P. intermedia/nigrescens, E. corrodens, F. nucleatum, and P. micros were the more prevalent pathogens in contrast to generalized AgP patients who harbored A. actinomycetemcomitans, P. gingivalis, and Capnocytophaga sp. as the most prevalent bacteria. CONCLUSIONS: C. rectus, P. gingivalis, E. corrodens, P. micros, and Capnocytophaga sp. were the most predominant periodontopathic bacteria of AgP in this Chilean population, but the only statistical difference found here between AgP and CP was for C. rectus, suggesting that the differences in clinical appearance may be caused by factors other than the microbiological composition of the subgingival plaque of these patients. In this study, the prevalence of A. actinomycetemcomitans was much lower than that of P. gingivalis.     

Total proteoglycan and chondroitin-4-sulfate levels in gingiva of patients with various types of periodontitis

BACKGROUND: The aim of the present study was to investigate the total proteoglycan (PG) and chondroitin-4-sulfate (C4S) levels in gingival tissue samples obtained from patients with aggressive periodontitis (AgP) and chronic periodontitis (CP) before therapy (baseline) and 1 month after completion of non-surgical periodontal therapy. METHODS: Gingival tissue samples were obtained from 10 AgP and 10 CP patients before initiation of treatment (baseline) and 1 month after non-surgical periodontal treatment. The control group comprised 10 systemically and periodontally healthy subjects. Total PG and C4S levels were determined by biochemical techniques. PG levels were analyzed using a modified Bitter and Muir method. C4S assay was carried out using chondroitin sulphate lyase AC and chondroitin-6 sulphate sulphohydrolase enzymes. The results were tested statistically using parametric tests. RESULTS: The clinical periodontal parameters demonstrated significant decreases in the periodontitis groups (P<0.05) after treatment, and there was no significant difference between AgP and CP groups at baseline and after treatment (P>0.05). At baseline, total PG and C4S levels in both of the periodontitis groups were significantly lower than that of the control group (P<0.05). One month after the non-surgical periodontal treatment, total PG levels in the periodontitis groups were comparable to the control group (P>0.05), whereas C4S levels in the AgP group were significantly lower than the other study groups (P<0.05). In the CP group, total PG and C4S levels increased significantly (P = 0.001 and P = 0.006, respectively) after non-surgical periodontal treatment, but they did not increase in the AgP group (P>0.05). CONCLUSION: The significant increases observed in total proteoglycan and chondroitin-4-sulfate levels after non-surgical periodontal treatment in the CP group but not in the AgP group may suggest that healing patterns differ between the two periodontitis types in terms of PG and C4S composition of extracellular matrix.     

Prevalence of periodontopathic bacteria in aggressive periodontitis patients in a Japanese population

BACKGROUND: Actinobacillus actinomycetemcomitans is considered a major etiologic agent of aggressive periodontitis. Other periodontopathic bacteria such as Porphyromonas gingivalis are also suspected of participating in aggressive periodontitis, although the evidence is controversial. The aim of the present study was to investigate the prevalence of periodontopathic bacteria and to clarify the microbiological features of aggressive periodontitis in Japanese patients. METHODS: Subgingival plaque was collected from 50 aggressive periodontitis (AgP) patients (localized 10, generalized 40). Samples from 35 generalized chronic periodontitis (CP) patients and 18 healthy subjects were examined as controls. Plaque samples were examined using culture and polymerase chain reaction (PCR) method. RESULTS: The prevalence of A. actinomycetemcomitans was relatively low in the localized (20%) and generalized (17.5%) AgP patients, with no significant difference observed in detection frequencies between AgP and the control groups (CP 8.6%, healthy 0%). On the other hand, Tannerella forsythensis (formerly Bacteroides forsythus), Campylobacter rectus, P. gingivalis, and Treponema denticola were frequently detected in localized as well as generalized aggressive periodontitis patients. The prevalence and proportion of P. gingivalis correlated with severity of clinical attachment loss in both localized and generalized aggressive periodontitis. CONCLUSIONS: T. forsythensis, C. rectus, P. gingivalis, and T. denticola were the predominant periodontopathic bacteria of aggressive periodontitis patients in Japan. Although A. actinomycetem- comitans was also detected in AgP patients, the prevalence of this bacterium was much lower than that of P. gingivalis.