BACKGROUND The quantitative alpha-defensin enzyme-linked immunosorbent assay(ELISA)demands a prior synovial fluid centrifugation,whereas this processing is not routinely required prior to the alpha-defensin lateral fl...BACKGROUND The quantitative alpha-defensin enzyme-linked immunosorbent assay(ELISA)demands a prior synovial fluid centrifugation,whereas this processing is not routinely required prior to the alpha-defensin lateral flow test.AIM To evaluate whether a prior synovial fluid centrifugation could lead the lateral flow performance to achieve comparable results to ELISA during periprosthetic joint infection(PJI)diagnosis.METHODS Fifty-three cases were included in this study:22 classified as PJI and 31 classified as aseptic cases,according to Musculoskeletal Infection Society 2013 criteria.Synovial fluid samples were submitted to centrifugation,and the supernatant was evaluated by ELISA and lateral flow tests.The sensitivity(SE),specificity(SP)and accuracy of each method were calculated as well as the agreement between those two methods.RESULTS In all of the 31 samples from aseptic patients,alpha-defensin ELISA and lateral flow tests showed negative results for infection.Regarding the 22 infected patients,the lateral flow test was positive in 19 cases(86.4%)and the ELISA was positive in 21(95.5%).Sensibility,SP and accuracy were,respectively,86.4%(95%CI:65.1%-97.1%),100%(95%CI:88.8%-100%)and 93.2%(95%CI:82.8%-98.3%)for the lateral flow test and 95.5%(95%CI:77.2%-99.9%),100%(95%CI:88.8%-100%)and 98.1%(95%CI:89.9%-100%)for ELISA.An agreement of 96.2%between those methods were observed.No statistical difference was found between them(P=0.48).CONCLUSION Alpha-defensin lateral flow test showed high SE,SP and accuracy after a prior synovial fluid centrifugation,achieving comparable results to ELISA.Considering the lower complexity of the lateral flow and its equivalent performance obtained in this condition,a prior centrifugation might be added as a valuable step to enhance the PJI diagnosis.展开更多
BACKGROUND The alpha-defensin lateral flow(ADLF)test is a new diagnostic tool for periprosthetic joint infection(PJI).Test accuracy for combined cohorts of hip and knee PJI has been reported to be good.AIM To assess t...BACKGROUND The alpha-defensin lateral flow(ADLF)test is a new diagnostic tool for periprosthetic joint infection(PJI).Test accuracy for combined cohorts of hip and knee PJI has been reported to be good.AIM To assess the accuracy of the ADLF test for hip PJI,and to compare three different diagnostic criteria for PJI.METHODS A cohort of 52 patients was identified,with a painful or poor-functioning total hip-or hemi-arthroplasty,that underwent aspiration and a subsequent ADLF test.PJI was diagnosed with Musculoskeletal Infection Society(MSIS)criteria,and sensitivity,specificity,overall accuracy,positive predictive value and negative predictive value were calculated.Furthermore,test specifics were compared with the European Bone and Joint Infection Society(EBJIS)and 2018 International Consensus Meeting(ICM)criteria for PJI.RESULTS Using MSIS criteria,sensitivity was 100%(CI:54%-100%)and specificity was 89%(CI:76%-96%).Six true positives and 5 false positives were found,including one case of metallosis.Using EBJIS criteria,more PJIs were found(11 vs 6),sensitivity was lower(71%,CI:42%-92%)and specificity was higher(97%,CI:86%-100%),with 4 false negatives and one false positive result.Using 2018 ICM criteria,sensitivity was 91%(62%-100%)and specificity 100%(91%-100%).The results in this cohort are comparable to previous studies.CONCLUSION Overall test accuracy of the ADLF test was good in this cohort,with a sensitivity of 100%and specificity of 89%.Using different PJI definition criteria,sensitivity and specificity changed slightly but overall accuracy remained around 90%.Using the ADLF test in metallosis cases can result in false positive results and should be performed with caution.展开更多
BACKGROUND Alpha-defensin has been widely studied for the diagnosis of periprosthetic joint infection(PJI).However,there is a lack of detailed information regarding the proper laboratory technique of the enzyme-linked...BACKGROUND Alpha-defensin has been widely studied for the diagnosis of periprosthetic joint infection(PJI).However,there is a lack of detailed information regarding the proper laboratory technique of the enzyme-linked immunosorbent assay(ELISA)method,such as sample dilution.AIM To assess the influence of dilution in the synovial fluid during ELISA for the diagnosis of knee PJI;and determine which dilution presents a better performance.METHODS Forty samples of synovial fluid from arthroplasty knees were included,17 in the infected group and 23 in the aseptic group,according to Musculoskeletal Infection Society criteria.Initially,five synovial fluid samples from each group were assessed for quantitative analysis of alpha-defensin using ELISA.Different dilution ratios(1:10,1:100,1:500,1:1000 and 1:5000)were tested based on the predetermined cutoff value of 5.2 mg/L.The dilutions that performed better were used to compare the results of all samples.RESULTS For infected cases,a gradual increase in the dilution of synovial fluid samples led to an equivalent increase in alpha-defensin level.The same was not observed in the aseptic cases.Both 1:1000 and 1:5000 dilutions presented satisfactory results to differentiate infected and aseptic cases.Further analyses were performed using 1:1000 and 1:5000 for all 40 samples.The 1:1000 dilution resulted in a sensitivity of 88.2%(95%CI,66%-98%)and specificity of 95.7%(95%CI,79%-99%),whereas the 1:5000 dilution presented a sensitivity of 94.1%(95%CI,73%-99%)and a specificity of 100%(95%CI,86%-100%).CONCLUSION The synovial fluid dilution had an important influence on the alpha-defensin ELISA results.Dilutions of 1:5000 showed the best performance for the diagnosis of knee PJI.The results of this study set the basis for a more reliable and reproducible alpha-defensin ELISA during the investigation of PJI,contributing to the expansion of this technique in different treatment centers worldwide.展开更多
背景:虽然关节置换围术期处置已经形成了一套标准化的流程,但是关节置换后假体周围感染仍是一个尚未彻底解决的难题,治疗方案依然没有达成共识。随着患者的不断增多,迫切需要具有指南意义的规范化治疗方案。目的:综述近年国外关节置换...背景:虽然关节置换围术期处置已经形成了一套标准化的流程,但是关节置换后假体周围感染仍是一个尚未彻底解决的难题,治疗方案依然没有达成共识。随着患者的不断增多,迫切需要具有指南意义的规范化治疗方案。目的:综述近年国外关节置换后假体周围感染的研究进展。方法:第一作者应用计算机检索1998至2019年PubMed数据库、Springerlink数据库的相关文章,英文检索词"Joint replacement,Periprosthetic joint infection,Progress in diagnosis and treatment";共检索到400余篇相关文献,50篇文献符合纳入标准。结果与结论:(1)随着新型血清标志物α-防御素和白细胞酯酶及D-二聚体、新型核医学检验方法及利用代谢组学和蛋白质组学等新方法的提出,可以凭借早期感染的生物标志物发现和诊断假体周围感染;(2)此外可以利用新的技术来破坏生物膜、微生物繁殖过程以及定量分子方法来提高病原体鉴定的准确性;(3)随着这些假体周围感染治疗进展的出现,快速、准确、经济的方法可能使明确诊断及培养出致病菌成为一件简单的事,那么很多病例都可以选择一期翻修;如若软组织条件不好或者不满足一期翻修的条件,则可进入二期翻修;(4)明确致病微生物,可以使用敏感的药物进行治疗,为二期翻修做好充足的准备,这对于假体周围感染治疗具有划时代的意义。展开更多
Objective: To evaluate the alpha-defensin (α-DF) genes polymorphism in women with vulvovaginal candidiasis and recurrence. Methods: This observational study included clinical vaginal secretion samples collected over ...Objective: To evaluate the alpha-defensin (α-DF) genes polymorphism in women with vulvovaginal candidiasis and recurrence. Methods: This observational study included clinical vaginal secretion samples collected over four years from 88 women, ranging in age from 18 to 65 years, from medical centers of Sao Paulo and Mogi das Cruzes, Brazil. Thirty-six of these women were asymptomatic (control group) and 52 presented clinical condition compatible with vulvovaginitis (38 primary or episodic as non-recurrent forms, and 14 recurrent vulvovaginal candidiasis). A portion of each sample was plated on Sabouraud dextrose agar with chloramphenicol and grown on CHROMagar Candida for presumptive characterization. The identification of the species was obtained by sequencing of the ITS1 region of rDNA. α-DF genes were amplified for subsequent evaluation of polymorphisms by endonuclease restriction assay. Results: From 88 samples were isolated 60 Candida albicans and 28 non-albicans Candida spp. Resistant C. albicans strains and non-albicans Candida spp. were more prevalent in recurrence. In all groups, the number of resistant non-albicans Candida spp. was most high than susceptible strains. α-DF1, α-DF3 and α-DF1/α-DF3 genotypes were found in 32 (36.4%), 17 (19.3%), 6 (6.8%) vaginal samples, respectively. About 33 samples were not amplified. Recurrence and severe disease were more observed in homozygous population. Conclusions: Non-albicans Candida spp. and homozygotic α-DF genotipes (α-DF1 and α-DF3) were more related with severe clinical signs and recurrence. Further studies about vulvovaginal candidiasis and α-DF genes are necessary to access the more comprehensive role of defensins in clinical manifestations.展开更多
基金Supported by Fundação de AmparoàPesquisa do Estado de São Paulo.
文摘BACKGROUND The quantitative alpha-defensin enzyme-linked immunosorbent assay(ELISA)demands a prior synovial fluid centrifugation,whereas this processing is not routinely required prior to the alpha-defensin lateral flow test.AIM To evaluate whether a prior synovial fluid centrifugation could lead the lateral flow performance to achieve comparable results to ELISA during periprosthetic joint infection(PJI)diagnosis.METHODS Fifty-three cases were included in this study:22 classified as PJI and 31 classified as aseptic cases,according to Musculoskeletal Infection Society 2013 criteria.Synovial fluid samples were submitted to centrifugation,and the supernatant was evaluated by ELISA and lateral flow tests.The sensitivity(SE),specificity(SP)and accuracy of each method were calculated as well as the agreement between those two methods.RESULTS In all of the 31 samples from aseptic patients,alpha-defensin ELISA and lateral flow tests showed negative results for infection.Regarding the 22 infected patients,the lateral flow test was positive in 19 cases(86.4%)and the ELISA was positive in 21(95.5%).Sensibility,SP and accuracy were,respectively,86.4%(95%CI:65.1%-97.1%),100%(95%CI:88.8%-100%)and 93.2%(95%CI:82.8%-98.3%)for the lateral flow test and 95.5%(95%CI:77.2%-99.9%),100%(95%CI:88.8%-100%)and 98.1%(95%CI:89.9%-100%)for ELISA.An agreement of 96.2%between those methods were observed.No statistical difference was found between them(P=0.48).CONCLUSION Alpha-defensin lateral flow test showed high SE,SP and accuracy after a prior synovial fluid centrifugation,achieving comparable results to ELISA.Considering the lower complexity of the lateral flow and its equivalent performance obtained in this condition,a prior centrifugation might be added as a valuable step to enhance the PJI diagnosis.
文摘BACKGROUND The alpha-defensin lateral flow(ADLF)test is a new diagnostic tool for periprosthetic joint infection(PJI).Test accuracy for combined cohorts of hip and knee PJI has been reported to be good.AIM To assess the accuracy of the ADLF test for hip PJI,and to compare three different diagnostic criteria for PJI.METHODS A cohort of 52 patients was identified,with a painful or poor-functioning total hip-or hemi-arthroplasty,that underwent aspiration and a subsequent ADLF test.PJI was diagnosed with Musculoskeletal Infection Society(MSIS)criteria,and sensitivity,specificity,overall accuracy,positive predictive value and negative predictive value were calculated.Furthermore,test specifics were compared with the European Bone and Joint Infection Society(EBJIS)and 2018 International Consensus Meeting(ICM)criteria for PJI.RESULTS Using MSIS criteria,sensitivity was 100%(CI:54%-100%)and specificity was 89%(CI:76%-96%).Six true positives and 5 false positives were found,including one case of metallosis.Using EBJIS criteria,more PJIs were found(11 vs 6),sensitivity was lower(71%,CI:42%-92%)and specificity was higher(97%,CI:86%-100%),with 4 false negatives and one false positive result.Using 2018 ICM criteria,sensitivity was 91%(62%-100%)and specificity 100%(91%-100%).The results in this cohort are comparable to previous studies.CONCLUSION Overall test accuracy of the ADLF test was good in this cohort,with a sensitivity of 100%and specificity of 89%.Using different PJI definition criteria,sensitivity and specificity changed slightly but overall accuracy remained around 90%.Using the ADLF test in metallosis cases can result in false positive results and should be performed with caution.
文摘BACKGROUND Alpha-defensin has been widely studied for the diagnosis of periprosthetic joint infection(PJI).However,there is a lack of detailed information regarding the proper laboratory technique of the enzyme-linked immunosorbent assay(ELISA)method,such as sample dilution.AIM To assess the influence of dilution in the synovial fluid during ELISA for the diagnosis of knee PJI;and determine which dilution presents a better performance.METHODS Forty samples of synovial fluid from arthroplasty knees were included,17 in the infected group and 23 in the aseptic group,according to Musculoskeletal Infection Society criteria.Initially,five synovial fluid samples from each group were assessed for quantitative analysis of alpha-defensin using ELISA.Different dilution ratios(1:10,1:100,1:500,1:1000 and 1:5000)were tested based on the predetermined cutoff value of 5.2 mg/L.The dilutions that performed better were used to compare the results of all samples.RESULTS For infected cases,a gradual increase in the dilution of synovial fluid samples led to an equivalent increase in alpha-defensin level.The same was not observed in the aseptic cases.Both 1:1000 and 1:5000 dilutions presented satisfactory results to differentiate infected and aseptic cases.Further analyses were performed using 1:1000 and 1:5000 for all 40 samples.The 1:1000 dilution resulted in a sensitivity of 88.2%(95%CI,66%-98%)and specificity of 95.7%(95%CI,79%-99%),whereas the 1:5000 dilution presented a sensitivity of 94.1%(95%CI,73%-99%)and a specificity of 100%(95%CI,86%-100%).CONCLUSION The synovial fluid dilution had an important influence on the alpha-defensin ELISA results.Dilutions of 1:5000 showed the best performance for the diagnosis of knee PJI.The results of this study set the basis for a more reliable and reproducible alpha-defensin ELISA during the investigation of PJI,contributing to the expansion of this technique in different treatment centers worldwide.
文摘背景:虽然关节置换围术期处置已经形成了一套标准化的流程,但是关节置换后假体周围感染仍是一个尚未彻底解决的难题,治疗方案依然没有达成共识。随着患者的不断增多,迫切需要具有指南意义的规范化治疗方案。目的:综述近年国外关节置换后假体周围感染的研究进展。方法:第一作者应用计算机检索1998至2019年PubMed数据库、Springerlink数据库的相关文章,英文检索词"Joint replacement,Periprosthetic joint infection,Progress in diagnosis and treatment";共检索到400余篇相关文献,50篇文献符合纳入标准。结果与结论:(1)随着新型血清标志物α-防御素和白细胞酯酶及D-二聚体、新型核医学检验方法及利用代谢组学和蛋白质组学等新方法的提出,可以凭借早期感染的生物标志物发现和诊断假体周围感染;(2)此外可以利用新的技术来破坏生物膜、微生物繁殖过程以及定量分子方法来提高病原体鉴定的准确性;(3)随着这些假体周围感染治疗进展的出现,快速、准确、经济的方法可能使明确诊断及培养出致病菌成为一件简单的事,那么很多病例都可以选择一期翻修;如若软组织条件不好或者不满足一期翻修的条件,则可进入二期翻修;(4)明确致病微生物,可以使用敏感的药物进行治疗,为二期翻修做好充足的准备,这对于假体周围感染治疗具有划时代的意义。
基金This study was financially supported by the Brazilian National Council for Scientific and Technological Development(CNPq,Brazil)We thank to Claudia R.C.Porto for support in laboratory tests.
文摘Objective: To evaluate the alpha-defensin (α-DF) genes polymorphism in women with vulvovaginal candidiasis and recurrence. Methods: This observational study included clinical vaginal secretion samples collected over four years from 88 women, ranging in age from 18 to 65 years, from medical centers of Sao Paulo and Mogi das Cruzes, Brazil. Thirty-six of these women were asymptomatic (control group) and 52 presented clinical condition compatible with vulvovaginitis (38 primary or episodic as non-recurrent forms, and 14 recurrent vulvovaginal candidiasis). A portion of each sample was plated on Sabouraud dextrose agar with chloramphenicol and grown on CHROMagar Candida for presumptive characterization. The identification of the species was obtained by sequencing of the ITS1 region of rDNA. α-DF genes were amplified for subsequent evaluation of polymorphisms by endonuclease restriction assay. Results: From 88 samples were isolated 60 Candida albicans and 28 non-albicans Candida spp. Resistant C. albicans strains and non-albicans Candida spp. were more prevalent in recurrence. In all groups, the number of resistant non-albicans Candida spp. was most high than susceptible strains. α-DF1, α-DF3 and α-DF1/α-DF3 genotypes were found in 32 (36.4%), 17 (19.3%), 6 (6.8%) vaginal samples, respectively. About 33 samples were not amplified. Recurrence and severe disease were more observed in homozygous population. Conclusions: Non-albicans Candida spp. and homozygotic α-DF genotipes (α-DF1 and α-DF3) were more related with severe clinical signs and recurrence. Further studies about vulvovaginal candidiasis and α-DF genes are necessary to access the more comprehensive role of defensins in clinical manifestations.
