Single And Combined Detection Of Anti-heart Autoantibodies In Diagnosis Of Viral Heart Disease

Part1Determination of autoantibodies against L-type calcium channel in patients with viral myocarditisObjective:Many studies showed that viral myocarditis (VMC) and dilated cardiomyopathy (DCM) were associated with viral infection and autoimmune reaction. So some scholars called them viral heart disease (VHD). Previous studies found that there were many anti-heart autoantibodies(AHA) in VHD patients, of which there were four kinds of AHA internationally recognized. They were anti-mitochondrial adenine nucleotide transfer enzyme antibody (anti-ANT), anti-betal adrenergic receptor antibody (anti-betalAR), anti M2-cholinergic receptor antibody (anti-M2) and anti-myosin heavy chain antibody (anti-MHC). The positive rate of these antibodies was higher in both VMC and DCM patients. They could be used as auxiliary diagnostic markers for VMC/DCM. Recently, Xiao Hua et al found a new autoantibodies against L-type calcium channel(CC-AAbs) in the blood of patients with DCM. The positive rate of CC-AAbs in the patients with DCM was significantly higher than that in normal control group. Then, was CC-AAbs also exist in VMC patients? How much was its positive rate? The purpose of this study was to detect CC-AAbs in patients with viral myocarditis and to evaluate the value of CC-AAbs in diagnosis of viral myocarditis.Methods:There were180individuals enrolled in this study, including acute viral myocarditis (AVMC) patients (60cases), DCM patients (60cases), and healthy blood donors (60cases). Enzyme-linked immunosorbent assay (ELISA) was used to detect CC-AAbs in the blood serum of three groups of subjects. The positive rate, sensitivity, specificity, and total agreement percentage were calculated.Result:The positive rate of CC-AAbs in AVMC group and DCM group were95%and96.7%. They were significantly higher than that(6.7%) in the healthy blood donors group(p<0.01). The sensitivity, specificity, and total agreement percentage of CC-AAbs in AVMC were95%,93.3%,94.2%. The sensitivity, specificity, and total agreement percentage of CC-AAbs in DCM were96.7%,93.3%,95%.Conclusion:There is also CC-AAbs in patients with AVMC. The positive rates of CC-AAbs in DCM and AVMC are significantly higher than that of healthy controls. The sensitivity, specificity, and total agreement percentage of CC-AAbs in DCM and AVMC are all higher. CC-AAbs can be used as an auxiliary diagnostic marker for VHD. Part2Single and combined detection of anti-heart autoantibodies in diagnosis of viral heart diseaseObjective:To develop and verify a kind of anti-heart autoantibodies(AHA) detection kit with high sensitivity and specificity. To investigate the diagnostic valuethe of single and combined detection of AHA in viral heart disease (VHD) patients.Methods:Myocardial mitochondrial adenine nucleotide translocase(ANT), β1adrenergic receptor(β1-AR), myosin heavy chain(MHC) and L-type calcium channel(L-CC) polypeptides with antigenic determinant were synthesized and produced as an anti-heart autoantibodies detection kit to detect serum AHA by enzyme-linked immunosorbent assay(ELISA). There are612individuals enrolled in this study, including acute viral myocarditis (AVMC) patients (78cases), dilated cardiomyopathy (DCM) patients (85cases), patients with other heart diseases and healthy individuals (a total of80cases). The sensitivity, specificity, and total agreement percentage were calculated. The data were analysed by the kappa in agreement.Result:The sensitivity, specificity, total agreement percentage of anti-ANT antibody in AVMC were93.6%,87.5%,90.5%, and compared with AVMC clinical diagnosis, kappa values was0.81, P<0.01. The sensitivity, specificity, total agreement percentage of anti-β1-AR antibody in AVMC were97.4%,75.0%,86.1%, and compared with AVMC clinical diagnosis, kappa values was0.72, P<0.01. The sensitivity, specificity, total agreement percentage of anti-MHC antibody in AVMC were87.2%,82.5%,84.8%, and compared with AVMC clinical diagnosis, kappa values was0.70, P<0.01. The sensitivity, specificity, total agreement percentage of anti-L-CC antibody in AVMC were97.4%,66.3%,81.0%, and compared with AVMC clinical diagnosis, kappa values was0.63, P<0.01. The sensitivity, specificity, total agreement percentage of anti-ANT antibody in DCM were85.9%,87.5%,86.7%, and compared with DCM clinical diagnosis, kappa values was0.73, P<0.01. The sensitivity, specificity, total agreement percentage of anti-β1-AR antibody in DCM were96.5%,75.0%,86.1%, and compared with DCM clinical diagnosis, kappa values was0.72, P<0.01. The sensitivity, specificity, total agreement percentage of anti-MHC antibody in DCM were92.9%,82.5%,87.9%, and compared with DCM clinical diagnosis, kappa values was0.76, P<0.0\. The sensitivity, specificity, total agreement percentage of anti-L-CC antibody in DCM were97.6%,66.3%,82.4%, and compared with DCM clinical diagnosis, kappa values was0.65, P<0.01. When the four antibodies were positive simultaneously, the sensitivity, specificity, total agreement percentage of four antibodies in diagnosis of AVMC were78.2%,92.5%,85.4%, and compared with AVMC clinical diagnosis, kappa value was0.71,p<0.01. When the four antibodies were positive simultaneously, the sensitivity, specificity, total agreement percentage of four antibodies in diagnosis of DCM were82.4%,92.5%,87.3%, and compared with AVMC clinical diagnosis, kappa value was0.75,p<0.01.Conclusion:The AHA results in AVMC and DCM through these kits we independently develop are highly consistent with clinical diagnosis. It can be used as an auxiliary diagnostic marker for AVMC and DCM. When detected alone, the value of anti-ANT in diagnosis of AVMC is the highest, the value of anti-MHC in diagnosis of DCM is the highest. Combined detection of four AHA can improve the specificity of diagnosis.