PBMC結(jié)核抗體分泌試驗(yàn)和酶聯(lián)免疫斑點(diǎn)試驗(yàn)對(duì)活動(dòng)性結(jié)核病臨床診斷價(jià)值的研究
發(fā)布時(shí)間：2019-06-20 11:09
【摘要】：目的：研究外周血單個(gè)核細(xì)胞（PBMC）分泌結(jié)核抗體試驗(yàn)和酶聯(lián)免疫斑點(diǎn)試驗(yàn)（ELISPOT）對(duì)活動(dòng)性結(jié)核�。═B）的臨床診斷價(jià)值。 方法：受試者共163例，包括活動(dòng)性結(jié)核病患者104例（菌陽(yáng)肺結(jié)核27例，菌陰肺結(jié)核61例，肺外結(jié)核16例）、結(jié)核分枝桿菌（MTB）潛伏感染者（LTBI）26例和健康對(duì)照（HC）33名。采用Ficoll密度梯度離心法分離受試者PBMCs，體外培養(yǎng)4d后收集培養(yǎng)上清液，用間接酶聯(lián)免疫吸附試驗(yàn)（ELISA）和蛋白免疫印跡法（WB）檢測(cè)PBMCs培養(yǎng)上清液中抗原特異性結(jié)核抗體；用ELISPOT方法檢測(cè)受試者早期分泌抗原靶6KDa蛋白（ESAT-6）、ESAT-6/培養(yǎng)濾出液蛋白-10（CFP-10）融合抗原刺激后產(chǎn)生γ-干擾素的T細(xì)胞水平；比較分析三種方法的敏感度、特異度、陽(yáng)性預(yù)測(cè)值及陰性預(yù)測(cè)值。 結(jié)果： 1. ELISA法檢測(cè)TB組特異性結(jié)核抗體的OD值高于LTBI組和HC組（p0.001）；特異性結(jié)核抗體區(qū)分TB和LTBI、HC的曲線下面積（AUC）分別為0.857、0.944，當(dāng)cut-off值為0.42時(shí)，區(qū)分TB和LTBI的敏感度為77.9%，，特異度為80.8%，陽(yáng)性預(yù)測(cè)值為94.2%，陰性預(yù)測(cè)值為47.7%；區(qū)分TB和HC的敏感度為77.9%，特異度為93.9%，陽(yáng)性預(yù)測(cè)值為97.6%，陰性預(yù)測(cè)值為57.4%。 2. WB法檢測(cè)活動(dòng)性結(jié)核的敏感度為79.2%，特異度為100.0%，陽(yáng)性預(yù)測(cè)值為100.0%，陰性預(yù)測(cè)值為40.7%（p0.001）。 3. ELISPOT法檢測(cè)活動(dòng)性結(jié)核的敏感度為82.7%，特異度為87.9%，陽(yáng)性預(yù)測(cè)值為95.6%，陰性預(yù)測(cè)值為61.7%（p0.001）。 4. ELISA、WB和ELISPOT方法診斷活動(dòng)性結(jié)核的敏感度、特異度、陽(yáng)性預(yù)測(cè)值進(jìn)行平行比較，差異均無(wú)統(tǒng)計(jì)學(xué)意義（p0.05）；陰性預(yù)測(cè)值進(jìn)行平行比較，WB法的陰性預(yù)測(cè)值低于ELISA法和ELISPOT法（p0.05）；ELISA和ELISPOT法在菌陽(yáng)組、菌陰組活動(dòng)性肺結(jié)核的陽(yáng)性檢出率比較，差異均無(wú)統(tǒng)計(jì)學(xué)意義（p0.05）。 5. ELISA、WB、ELISOPT法檢測(cè)16例肺外結(jié)核的陽(yáng)性率分別為81.3%、56.3%、68.8%。 結(jié)論： 1. PBMC結(jié)核抗體分泌試驗(yàn)的ELISA法，對(duì)活動(dòng)性結(jié)核的診斷效能高，并可以區(qū)分活動(dòng)性結(jié)核、結(jié)核分枝桿菌潛伏感染及健康人。 2. PBMC結(jié)核抗體分泌試驗(yàn)的免疫印跡法操作簡(jiǎn)便，診斷活動(dòng)性結(jié)核的特異度高。 3.PBMC結(jié)核抗體分泌試驗(yàn)與ELISPOT法對(duì)活動(dòng)性結(jié)核均具有較高的診斷價(jià)值，PBMC結(jié)核抗體分泌試驗(yàn)可以作為結(jié)核病實(shí)驗(yàn)室診斷的新方法。
[Abstract]:Objective: to study the clinical diagnostic value of tuberculosis antibody test and enzyme-linked immunosorbent assay (ELISPOT) secreted by peripheral blood mononuclear cells (PBMC) in active tuberculosis (TB). Methods: a total of 163 subjects, including 104 cases of active tuberculosis (27 cases of positive pulmonary tuberculosis, 61 cases of negative pulmonary tuberculosis, 16 cases of extrapulmonary tuberculosis), 26 cases of latent infection of Mycobacterium tuberculosis (MTB) and 33 cases of healthy control (HC). The culture medium of PBMCs, was isolated by Ficoll density gradient centrifugation for 4 days, and the antigen-specific tuberculosis antibody in the culture medium of PBMCs was detected by indirect enzyme-linked immunosorbent assay (ELISA) and Western immunoblotting (WB). ELISPOT method was used to detect the T cell level of interferon gamma induced by early secretion of antigen target 6KDa protein (ESAT-6) and ESAT-6/ culture filtrate protein 10 (CFP-10) fusion antigen, and the sensitivity, specificity, positive predictive value and negative predictive value of the three methods were compared and analyzed. Results: 1. The OD value of specific tuberculosis antibody in TB group was higher than that in LTBI group and HC group (p0.001), and the area (AUC) under the curve of specific tuberculosis antibody distinguishing TB and LTBI,HC was 0.857 and 0.944, respectively. when cut-off value was 0.42, the sensitivity, specificity, positive predictive value and negative predictive value of distinguishing TB and LTBI were 77.9%, 80.8%, 94.2% and 47.7%, respectively. when cut-off value was 0.42%, the sensitivity, specificity, positive predictive value and negative predictive value of TB and LTBI were 77.9%, 80.8%, 94.2% and 47.7%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of TB and HC were 77.9%, 93.9%, 97.6% and 57.4%, respectively. 2. The sensitivity, specificity, positive predictive value and negative predictive value of WB for the detection of active tuberculosis were 79.2%, 100.0%, 100.0% and 40.7% (p0.001) respectively. 3. The sensitivity, specificity, positive predictive value and negative predictive value of ELISPOT in detecting active tuberculosis were 82.7%, 87.9%, 95.6% and 61.7% (p0.001) respectively. 4. There was no significant difference in the sensitivity, specificity and positive predictive value of ELISA,WB and ELISPOT in the diagnosis of active tuberculosis (p0.05), but the negative predictive value of WB was lower than that of Elisa and ELISPOT (p0.05); ELISA and ELISPOT). There was no significant difference in the positive rate of active pulmonary tuberculosis between the negative group and the negative group (p0.05). 5. The positive rates of 16 cases of extrapulmonary tuberculosis detected by ELISA,WB,ELISOPT were 81.3%, 56.3% and 68.8%, respectively. Conclusion: 1. ELISA method of PBMC tuberculosis antibody secretion test is effective in the diagnosis of active tuberculosis, and can distinguish active tuberculosis, latent infection of Mycobacterium tuberculosis and healthy people. 2. The immunoblotting method of PBMC tuberculosis antibody secretion test is simple and has high specificity in the diagnosis of active tuberculosis. 3.PBMC tuberculosis antibody secretion test and ELISPOT method have high diagnostic value for active tuberculosis. PBMC tuberculosis antibody secretion test can be used as a new method for laboratory diagnosis of tuberculosis.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R52
本文編號(hào)：2503173
[Abstract]:Objective: to study the clinical diagnostic value of tuberculosis antibody test and enzyme-linked immunosorbent assay (ELISPOT) secreted by peripheral blood mononuclear cells (PBMC) in active tuberculosis (TB). Methods: a total of 163 subjects, including 104 cases of active tuberculosis (27 cases of positive pulmonary tuberculosis, 61 cases of negative pulmonary tuberculosis, 16 cases of extrapulmonary tuberculosis), 26 cases of latent infection of Mycobacterium tuberculosis (MTB) and 33 cases of healthy control (HC). The culture medium of PBMCs, was isolated by Ficoll density gradient centrifugation for 4 days, and the antigen-specific tuberculosis antibody in the culture medium of PBMCs was detected by indirect enzyme-linked immunosorbent assay (ELISA) and Western immunoblotting (WB). ELISPOT method was used to detect the T cell level of interferon gamma induced by early secretion of antigen target 6KDa protein (ESAT-6) and ESAT-6/ culture filtrate protein 10 (CFP-10) fusion antigen, and the sensitivity, specificity, positive predictive value and negative predictive value of the three methods were compared and analyzed. Results: 1. The OD value of specific tuberculosis antibody in TB group was higher than that in LTBI group and HC group (p0.001), and the area (AUC) under the curve of specific tuberculosis antibody distinguishing TB and LTBI,HC was 0.857 and 0.944, respectively. when cut-off value was 0.42, the sensitivity, specificity, positive predictive value and negative predictive value of distinguishing TB and LTBI were 77.9%, 80.8%, 94.2% and 47.7%, respectively. when cut-off value was 0.42%, the sensitivity, specificity, positive predictive value and negative predictive value of TB and LTBI were 77.9%, 80.8%, 94.2% and 47.7%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of TB and HC were 77.9%, 93.9%, 97.6% and 57.4%, respectively. 2. The sensitivity, specificity, positive predictive value and negative predictive value of WB for the detection of active tuberculosis were 79.2%, 100.0%, 100.0% and 40.7% (p0.001) respectively. 3. The sensitivity, specificity, positive predictive value and negative predictive value of ELISPOT in detecting active tuberculosis were 82.7%, 87.9%, 95.6% and 61.7% (p0.001) respectively. 4. There was no significant difference in the sensitivity, specificity and positive predictive value of ELISA,WB and ELISPOT in the diagnosis of active tuberculosis (p0.05), but the negative predictive value of WB was lower than that of Elisa and ELISPOT (p0.05); ELISA and ELISPOT). There was no significant difference in the positive rate of active pulmonary tuberculosis between the negative group and the negative group (p0.05). 5. The positive rates of 16 cases of extrapulmonary tuberculosis detected by ELISA,WB,ELISOPT were 81.3%, 56.3% and 68.8%, respectively. Conclusion: 1. ELISA method of PBMC tuberculosis antibody secretion test is effective in the diagnosis of active tuberculosis, and can distinguish active tuberculosis, latent infection of Mycobacterium tuberculosis and healthy people. 2. The immunoblotting method of PBMC tuberculosis antibody secretion test is simple and has high specificity in the diagnosis of active tuberculosis. 3.PBMC tuberculosis antibody secretion test and ELISPOT method have high diagnostic value for active tuberculosis. PBMC tuberculosis antibody secretion test can be used as a new method for laboratory diagnosis of tuberculosis.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R52
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本文編號(hào)：2503173
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