【摘要】：第一部分病毒性心肌炎患者抗鈣通道抗體的測定 目的：眾多研究表明,病毒性心肌炎(VMC)、擴張型心肌病(DCM)均與病毒感染及其后的自身免疫反應(yīng)有關(guān),故有學(xué)者將二者統(tǒng)稱為病毒性心臟病(VHD)。既往研究發(fā)現(xiàn),在VHD患者體內(nèi)存在多種抗心肌自身抗體(AHA),其中國際上較為公認的四種抗心肌肽類抗體是抗線粒體腺嘌呤核苷酸轉(zhuǎn)移酶抗體(anti-ANT)、抗β1腎上腺素能受體抗體(anti-β1AR)、抗M2-膽堿能受體抗體(anti-M2)和抗肌球蛋白重鏈抗體(anti-MHC)。這些抗體在VMC和DCM患者中均有較高的陽性檢出率,可用于輔助診斷VMC和DCM。最近,肖華等發(fā)現(xiàn)在DCM患者的血液中存在一種新的抗L型鈣離子通道自身抗體(CC-AAbs),該抗體在DCM患者中的陽性檢出率顯著高于正常對照組。那么在VMC患者體內(nèi)是否也存在著這種抗L型鈣通道自身抗體呢?其陽性率又如何?本研究旨在檢測VMC患者血液中的抗L型鈣通道抗體,探討其對病毒性心肌炎的診斷價值。 方法：收集在華中科技大學(xué)同濟醫(yī)學(xué)院附屬協(xié)和醫(yī)院心血管內(nèi)科就診和(或)住院的急性病毒性心肌炎(AVMC)、DCM患者各60例,并以60例健康獻血者作為對照,采用酶聯(lián)免疫吸附法(ELISA)檢測三組受試者血清中的CC-AAbs,計算陽性率、靈敏度、特異度、符合率。 結(jié)果：AVMC組、DCM組CC-AAbs的陽性率分別為95%、96.7%,明顯高于健康對照組的6.7%(p值均0.01),CC-AAbs對AVMC檢測的靈敏度、特異度與符合率分別為95%、93.3%、94.2%,CC-AAbs對DCM檢測的靈敏度、特異度與符合率分別為96.7%、93.3%、95%。 結(jié)論：AVMC患者體內(nèi)也存在CC-AAbs。CC-AAbs在AVMC、DCM中的陽性率顯著高于健康對照組。CC-AAbs對AVMC、DCM檢測的靈敏度、特異度與符合率均較高,可用于輔助診斷病毒性心臟病。 第二部分單獨和聯(lián)合檢測抗心肌抗體診斷病毒性心臟病 目的：開發(fā)并驗證一種高靈敏度和高特異度的抗心肌抗體(AHA)檢測試劑盒,探討單獨和聯(lián)合檢測AHA對病毒性心臟病(VHD)的診斷價值。 方法：合成具有抗原決定簇的心肌線粒體腺嘌呤核苷酸轉(zhuǎn)運體(／ANT)、β1腎上腺素能受體(p1-AR)、肌球蛋白重鏈(MHC)、L-型鈣離子通道(L-CC)的多肽,制成相應(yīng)的AHA檢測試劑盒,用酶聯(lián)免疫吸附法(ELISA)檢測急性病毒性心肌炎(AVMC)患者(78例)、擴張型心肌病(DCM)患者(85例)、其他心臟病患者及健康體檢者(共80例)血清中的四種AHA,以臨床診斷參考標準為金標準,計算這四種AHA診斷AVMC、DCM的靈敏度、特異度、符合率,并行kappa一致性檢驗。 結(jié)果：抗ANT抗體診斷AVMC的靈敏度、特異度、符合率分別為93.6%、87.5%、90.5%,與臨床診斷比較,kappa值為0.81,P值0.01；抗β1-AR抗體診斷AVMC的靈敏度、特異度、符合率分別為97.4%、75.0%、86.1%,與臨床診斷比較,kappa值為0.72,P值0.01；抗MHC抗體診斷AVMC的靈敏度、特異度、符合率分別為87.2%、82.5%、84.8%,與臨床診斷比較,kappa值為0.70,P值0.01；抗L-CC抗體診斷AVMC的靈敏度、特異度、符合率分別為97.4%、66.3%、81.0%,與臨床診斷比較,kappa值為0.63,P值0.01。抗ANT抗體診斷DCM的靈敏度、特異度、符合率分別為85.9%、87.5%、86.7%,與臨床診斷比較,kappa值為0.73,P值0.01；抗β1-AR抗體診斷DCM的靈敏度、特異度、符合率分別為96.5%、75.0%、86.1%,與臨床診斷比較,kappa值為0.72,P值0.01；抗MHC抗體診斷DCM的靈敏度、特異度、符合率分別為92.9%、82.5%、87.9%,與臨床診斷比較,kappa值為0.76,P值0.01；抗L-CC抗體診斷DCM的靈敏度、特異度、符合率分別為97.6%、66.3%、82.4%,與臨床診斷比較,kappa值為0.65,P值0.01。若同一份血清四種AHA均陽性診斷AVMC的靈敏度、特異度、符合率分別為78.2%、92.5%、85.4%,與臨床診斷比較,kappa值為0.71,P值0.01。同一份血清四種AHA均陽性診斷DCM的靈敏度、特異度、符合率分別為82.4%、92.5%、87.3%,與臨床診斷比較,kappa值為0.75,P值0.01。 結(jié)論：我們自主研發(fā)的系列AHA試劑盒檢測AVMC和DCM的結(jié)果與其臨床診斷具有高度一致性,可作為VHD的輔助診斷指標。單獨檢測時anti-ANT對AVMC的診斷價值最高,anti-MHC對DCM的診斷價值最高,四種抗體聯(lián)合檢測能提高診斷的特異度。
[Abstract]:Part 1 Determination of anti calcium channel antibody in patients with viral myocarditis
Objective: Many studies have shown that viral myocarditis (VMC) and dilated cardiomyopathy (DCM) are related to viral infection and subsequent autoimmune response, so some scholars call them viral heart disease (VHD). Previous studies have found that there are many kinds of anti-myocardial autoantibodies (AHA) in patients with VHD, of which four are internationally recognized anti-myocardial autoantibodies. Cardiac peptide antibodies are anti-mitochondrial adenine nucleotide transferase antibodies (anti-ANT), anti-beta-1-adrenergic receptor antibodies (anti-beta 1AR), anti-M2-cholinergic receptor antibodies (anti-M2) and anti-myosin heavy chain antibodies (anti-MHC). These antibodies have high positive detection rates in VMC and DCM patients and can be used to assist the diagnosis of VMC and DCM. Recently, Xiao Hua et al. found a new anti-L-type calcium channel autoantibody (CC-AAbs) in the blood of patients with DCM. The positive rate of CC-AAbs in patients with DCM was significantly higher than that in normal control group. Objective to investigate the diagnostic value of anti L type calcium channel antibody in patients with MC.
Methods: 60 patients with acute viral myocarditis (AVMC) and 60 patients with DCM were enrolled in the Department of Cardiovascular Medicine, Union Medical College Affiliated to Tongji Medical College, Huazhong University of Science and Technology, and 60 healthy blood donors were used as control group. The serum CC-AAbs were detected by enzyme-linked immunosorbent assay (ELISA). Dissimilarity, coincidence rate.
Results: The positive rates of CC-AAbs in AVMC group and DCM group were 95%, 96.7%, significantly higher than 6.7% in healthy control group (p 0.01). The sensitivity, specificity and coincidence rate of CC-AAbs to AVMC were 95%, 93.3%, 94.2% respectively. The specificity and coincidence rate of CC-AAbs to DCM were 96.7%, 93.3% and 95% respectively.
Conclusion: The positive rate of CC-AAbs. CC-AAbs in AVMC patients is significantly higher than that in healthy controls. CC-AAbs has higher sensitivity, specificity and coincidence rate in detecting AVMC and DCM, and can be used as an auxiliary diagnostic tool for viral heart disease.
The second part is the separate and joint detection of anti myocardial antibodies in the diagnosis of viral heart disease.
Objective: To develop and validate a high sensitivity and specificity test kit for detecting anti-myocardial antibody (AHA), and to explore the diagnostic value of single and combined detection of AHA for viral heart disease (VHD).
METHODS: Polypeptides of adenine nucleotide transporter (/ANT), beta-1-adrenergic receptor (p1-AR), myosin heavy chain (MHC), L-type calcium channel (L-CC) with antigenic determinants were synthesized, and the corresponding AHA kits were made. 78 patients with acute viral myocarditis (AVMC) were detected by enzyme-linked immunosorbent assay (ELISA). Four kinds of AHA in serum of 85 patients with dilated cardiomyopathy (DCM), 80 patients with other heart diseases and 80 healthy persons were calculated according to the clinical diagnostic reference standard. The sensitivity, specificity and coincidence rate of the four AHA in diagnosis of AVMC and DCM were calculated and the kappa consistency test was performed.
Results: The sensitivity and specificity of anti-ANT antibody in the diagnosis of AVMC were 93.6%, 87.5% and 90.5%, respectively. Compared with clinical diagnosis, the kappa value was 0.81, P value was 0.01; the sensitivity and specificity of anti-beta 1-AR antibody in the diagnosis of AVMC were 97.4%, 75.0%, 86.1%, respectively. The sensitivity, specificity and coincidence rate were 87.2%, 82.5% and 84.8% respectively. Compared with clinical diagnosis, the kappa value was 0.70 and P value was 0.01. The sensitivity, specificity and coincidence rate of anti-L-CC antibody in diagnosis of AVMC were 97.4%, 66.3% and 81.0% respectively. Compared with clinical diagnosis, the kappa value was 0.63 and P value was 0.01. Compared with clinical diagnosis, the kappa value was 0.73, P value was 0.01; the sensitivity and specificity of anti-beta-1-AR antibody were 96.5%, 75.0%, 86.1%, respectively. Compared with clinical diagnosis, the kappa value was 0.72, P value was 0.01; the sensitivity and specificity of anti-MHC antibody in diagnosis of DCM were 92.9%, 82.5%, 87.9%, respectively. The sensitivity and specificity of anti-L-CC antibody were 97.6%, 66.3% and 82.4%, respectively. Compared with clinical diagnosis, the kappa value was 0.65 and the P value was 0.01. The sensitivity, specificity and coincidence rate of four kinds of AHA in the same serum were 82.4%, 92.5% and 87.3% respectively. Compared with clinical diagnosis, the kappa value was 0.75 and P value was 0.01.
Conclusion: The results of our series of AHA kits for detecting AVMC and DCM are highly consistent with their clinical diagnosis and can be used as an auxiliary diagnostic index for VHD.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R541

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