Davydova J, Pankuweit S, Crombach M, Eckhardt H, Strache D, Faulhammer P, Maisch B
Moscow Setschenov Academy, Russia.
Herz. 2000 May;25(3):233-9. doi: 10.1007/s000590050012.
The development of highly sensitive molecular biological methods such as in-situ hybridization and polymerase chain reaction (PCR) made it possible to detect viral/bacterial nucleic acid in human endomyocardial biopsies. However, only a few investigations addressed the problem of latent persistence of viral and bacterial genome and the detection of the corresponding proteins, which could have important consequences for the clinical course of the disease. The purpose of this study was to determine whether protein of various viruses (adenovirus, enterovirus, cytomegalovirus, influenza A and B virus, herpes simplex virus 1 and 2) and bacteria (chlamydia pneumonia) can be detected in endomyocardial biopsies of patients with myocarditis and dilated cardiomyopathy with and without inflammation by use of an immunofluorescence assay and to compare the frequency of its detection with the results of PCR, immunohistology and serology. Thirty-nine patients with myocarditis and dilated cardiomyopathy with and without inflammation were examined by a direct immunofluorescence assay using the endomyocardial biopsy as antigen. Each of the samples was additionally studied by immunohistological methods and PCR for the detection of infiltrating cells and the genome of cardiotropic viruses or bacteria. Fourteen of patients were considered to have myocarditis (group 1), 9 dilated cardiomyopathy with inflammation (group 2), 10 dilated cardiomyopathy (group 3), 6 to have no myocarditis or dilated cardiomyopathy (group 4). Using a direct immunofluorescence assay we could show only that 1 patient without histological myocarditis or dilated cardiomyopathy (group 4) was positive for influenza B and chlamydia pneumonia antigens in the endomyocardial biopsy. In addition we have determined influenza B-specific antibodies, such as IgG (marginal titer) and IgA (high titer) and chlamydia pneumonia-specific antibodies, such as IgG (marginal titer) in serum of this patient. A second patient with dilated cardiomyopathy was found to be positive for protein of chlamydia pneumonia, who was shown to have chlamydia pneumonia-specific antibodies, such as IgG (high titer) in serum. There was no correlation with PCR results, but good correlation with influenza B and chlamydia pneumonia-specific antibodies in sera of these patients. In this investigation we have determined viral/bacterial-specific antibodies using serological methods and proteins of these agents using immunoflourescence. Despite the detection of virus or bacteria-specific antibodies in the sera and detection of viral and/or bacterial protein in the biopsies of some of the patients viral and/or bacterial genome was not found in the biopsy. This may be explained by the focal character of myocarditis and sampling error, because for technical reasons we use different biopsies for immunohistochemical and molecular biological investigations.
高灵敏度分子生物学方法如原位杂交和聚合酶链反应（PCR）的发展，使得在人类心内膜心肌活检中检测病毒/细菌核酸成为可能。然而，仅有少数研究探讨了病毒和细菌基因组的潜伏持续性问题以及相应蛋白质的检测，而这可能对疾病的临床病程产生重要影响。本研究的目的是确定在有炎症和无炎症的心肌炎及扩张型心肌病患者的心内膜心肌活检中，是否能够通过免疫荧光测定法检测到多种病毒（腺病毒、肠道病毒、巨细胞病毒、甲型和乙型流感病毒、单纯疱疹病毒1型和2型）和细菌（肺炎衣原体）的蛋白质，并将其检测频率与PCR、免疫组织学和血清学结果进行比较。39例有炎症和无炎症的心肌炎及扩张型心肌病患者接受了以心内膜心肌活检为抗原的直接免疫荧光测定。每个样本还通过免疫组织学方法和PCR进行研究，以检测浸润细胞以及嗜心性病毒或细菌的基因组。14例患者被认为患有心肌炎（第1组），9例患有炎症性扩张型心肌病（第2组），10例患有扩张型心肌病（第3组），6例无心肌炎或扩张型心肌病（第4组）。通过直接免疫荧光测定，我们仅发现1例无组织学心肌炎或扩张型心肌病的患者（第4组）在心内膜心肌活检中乙型流感病毒和肺炎衣原体抗原呈阳性。此外，我们还测定了该患者血清中的乙型流感病毒特异性抗体，如IgG（临界滴度）和IgA（高滴度），以及肺炎衣原体特异性抗体，如IgG（临界滴度）。另1例扩张型心肌病患者被发现肺炎衣原体蛋白呈阳性，其血清中显示有肺炎衣原体特异性抗体，如IgG（高滴度）。与PCR结果无相关性，但与这些患者血清中的乙型流感病毒和肺炎衣原体特异性抗体有良好相关性。在本研究中，我们使用血清学方法测定了病毒/细菌特异性抗体，并使用免疫荧光法测定了这些病原体的蛋白质。尽管在一些患者的血清中检测到了病毒或细菌特异性抗体，且在活检中检测到了病毒和/或细菌蛋白，但在活检中未发现病毒和/或细菌基因组。这可能是由于心肌炎的局灶性特征和采样误差所致，因为出于技术原因，我们在免疫组织化学和分子生物学研究中使用了不同的活检样本。
