二代测序在膝关节置换术后迟发性感染关节液中病原菌检测中的应用价值

Application value of next-generation sequencing technology in the detection of pathogenic bacteria in the joint fluid of delayed infection after knee arthroplasty

目的探讨二代测序在膝关节置换术后迟发性感染关节液中病原菌检测中的应用价值。方法2014年2月~2019年3月我院收治的膝关节置换术后合并感染病人41例。收集所有病人关节液,对关节液中细菌分别进行二代测序及传统细菌培养鉴定。统计两种方法对不同细菌的检出率。结果二代测序检出感染革兰阳性菌金黄色葡萄球菌的病人31.71%、表皮葡萄球菌21.95%、溶血葡萄球菌24.39%、人葡萄球菌14.63%、无乳链球菌17.07%、化脓性链球菌9.76%、停乳链球菌12.20%;感染革兰阴性菌铜绿假单胞菌的病人19.51%、大肠埃希菌14.63%、流感嗜血杆菌4.88%、肺炎克雷伯菌12.20%、鲍氏不动杆菌14.63%。细菌培养未检出停乳链球菌,两种方法均未检出真菌。二代测序检出的感染金黄色葡萄球菌病人13例(70.73%,13/41)明显多于细菌培养5例(12.20%,5/41)(P<0.05)。二代测序得到41例病人样本中细菌平均ACE指数(12834.06±317.52),平均Chao1指数(8025.37±351.08),Simpson指数(0.53±0.01)、Shannon指数(5.06±0.11)。结论二代测序在对膝关节置换术后感染的病人关节液中细菌检出的全面性上具有较大优势,应用前景及价值均较好。

Objective To investigate the application value of next-generation sequencing technology in the detection of pathogenic bacteria in the joint fluid of delayed infection after knee arthroplasty.Methods 41 patients with joint infection after knee replacement in our hospital from February 2014 to March 2019 were selected.The bacteria in joint fluid were identified by next-generation sequencing technology and traditional bacterial culture.Then the detection rate of the two methods for different bacteria was calculated.Results In the second-generation sequencing,31.71%,21.95%,24.39%,14.63%,17.07%,9.76% and 12.20% of the patients infected with gram-positive Staphylococcus aureus,Staphylococcus epidermidis,Staphylococcus haemolyticus,human Staphylococcus,Streptococcus lactis,Streptococcus pyogenes and Streptococcus lactis were detected,moreover,19.51%,14.63%,4.88%,12.20% and 14.63% of the patients were infected with gram-negative Pseudomonas aeruginosa,Escherichia coli,Haemophilus influenzae,Klebsiella pneumoniae and Acinetobacter baumannii.Bacterial culture found no Streptococcus lactis.No fungi were detected by the two methods.The detection rate of Staphylococcus aureus was 70.73%(13/41) by next-generation sequencing technology,which was higher than 12.20%(5/41) by traditional bacteria culture(P<0.05).The next-generation sequencing technology results showed that the average bacterial ACE index(12 834.06±317.52),Chao1 index(8 025.37±351.08),Simpson index(0.53±0.01) and Shannon index(5.06±0.11) in the samples of 41 patients were obtained.Conclusion next-generation sequencing technology has a great advantage in the detection of bacteria in the joint fluid of infected patients after knee replacement,has good application prospect and value.