难治性根尖周炎患者HK1 mRNA检测、感染病原菌分布及牙龈卟啉单胞菌基因型分析

HK1 detection, distribution of pathogenic bacteria, and Porphyromonas gingivalis gene detection in patients with refractory periapical periodontitis

目的检测难治性根尖周炎患者血糖激酶1(HK1)水平,并分析感染病原菌分布及牙龈卟啉单胞菌基因型,为该病的防治提供参考。方法选择2018年6月-2019年6月本院收治的难治性根尖周炎患者53例(病例组),正常健康者50例(对照组),采用荧光定量PCR检测临床标本中的HK1 mRNA水平,并进行比较分析;培养法检测患者感染病原菌种类,PCR法检测感染牙龈卟啉单胞菌的基因型,K-B纸片扩散法检测病原菌的耐药性。结果与对照组比较,难治性根尖周炎患者临床标本中的HK1 mRNA高表达(P<0.05)。53例患者共检出85株病原菌,其中革兰阳性菌50株(58.82%),主要为葡萄球菌属(21.18%)、粪肠球菌(12.94%)和微单胞菌(11.76%);革兰阴性菌35株(41.18%),以牙龈卟啉单胞菌(27.06%)为主。葡萄菌属、粪肠球菌以及微单胞菌对美罗培南、亚胺培南、头孢曲松的耐药率均<50.00%,牙龈卟啉单胞菌对氯霉素、头孢菌素类和庆大霉素等的耐药率均>50.00%。23株牙龈卟啉单胞菌基因型占比分别为:fimAⅡ型34.78%,fimAⅠ型21.75%,kgpⅠ型56.52%,kgpⅡ型43.48%。常见fimA和kgp基因型组合为fimAⅡ型+kgpⅠ型和fimAⅡ型+kgpⅡ型,占17.39%(4/23)。肿胀患者感染的牙龈卟啉单胞菌以fimAⅡ型(40.00%)和kgpⅠ型(60.00%)为主,窦道患者以fimAⅡ型(46.15%)感染为主,kgpⅠ型与kgpⅡ感染占比相近。结论难治性根尖周炎感染根管感染病原菌以革兰阳性菌为主,不同基因型牙龈卟啉单胞菌引起疾病的临床表现可能不同。患者血HK1高表达,其在难治性根尖周炎中的作用有待进一步研究。

Objective To detect the level of glucose kinase 1(HK1)in patients with refractory periapical periodontitis,and to analyze the distribution of pathogenic bacteria and Porphyromonas gingivalis genotypes,in order to provide a reference for the prevention and treatment of the disease.Methods patients with refractory periapical periodontitis seen at this hospital from June 2018 to June 2019 served as the patient group,and 50 healthy people served as the control group.The levels of HK1 mRNA in clinical samples were detected using fluorescence quantitative PCR and compared.The species of pathogenic bacteria were detected by culture.PCR was used to detect the genotype of P.gingivalis.The K-B disk diffusion method was used to determinet the drug resistance of pathogenic bacterias.Results The levels of HK1 mRNA expression in patients was significantly higher than that in the control group(P<0.05).Eighty-five strains of pathogenic bacteria were detected in 53 patients.Fifty(58.82%)of those were strains of Gram-positive bacteria,which were mainly Staphylococcus(21.18%),Enterococcus faecalis(12.94%)and Parvimonas(11.76%).There were 35(41.18%)strains of Gram-negative bacteria,of which P.gingivalis(27.06%)was predominant.The resistance of Staphylococcus,Enterococcus faecalis,and Parvimonas to meropenem,imipenem,and ceftriaxone was lower than 50.00%,while the resistance of P.gingivalis to chloramphenicol,cephalosporins,and gentamicin were higher than 50.00%.In 23 samples of P.gingivalis,fimA II was detected at a rate of 34.71%,fimA I was detected at a rate of 21.75%,kgp I was detected at a rate of 56.52%,and kgp II was detected at a rate of 43.48%.The common genotype combinations of fimA and kgp were fimAII+kgpI and fimAII+kgpII,accounting for 17.39%(4/23).fimAII(40.00%)and kgpI(60.00%)were the main pathogens in patients with swelling.fimAII(46.15%)was the main type of infection in patients with a sinus tract,and the proportion of kgpI and kgpII was similar.Conclusion Gram-positive bacteria were the main pathogens causing a root canal infection in refractory periapical periodontitis.Different genotypes of P.gingivalis may cause different clinical manifestations.The high level of HK1 expression in patients with refractory periapical periodontitis needs to be studied further.