柯萨奇B组病毒IgM抗体特性研究

目的 研究柯萨奇B组病毒（CVB）感染后患者急性期CVB－IgM抗体的反应特性。方法 对临床确诊为CVB感染的患者急性期血清用免疫印迹法检测CVB－IgM抗体。结果 患者的急性期血清IgM抗体均仅针对CVB的VP1抗原反应。经ELISA与免疫印迹方法的比较表明：用免疫印迹法检测的CVB－IgM抗体可对CVB各型病毒抗原反应,具有CVB各型病毒之间的交 叉反应性。而同时,在16例ELISA检测反应阴性的健康人血清中未见有CVB的VP1特异性的IgM抗体存在。结论 临床感染CVB的患者急性期,其血清中的特异性的IgM抗体是针对CVB的VP1抗原,而且,此类抗体对各型的CVB抗原具有交叉反应性。     

Enzyme-linked immunosorbent assay for the detection and identification of coxsackie B antigen in tissue cultures and clinical specimens

An enzyme-linked immunosorbent assay (ELISA) has been developed for the identification of Coxsackie B antigens. This assay was capable of identifying and distinguishing all six Coxsackie B serotypes at concentrations one hundredfold to ten thousandfold less than could be detected by complement fixation (CF) systems. In addition, the Coxsackie B ELISA correctly identified the presence of Coxsackie B antigen in 19 of 21 tissue culture fluids and five of nine rectal swab specimens. Two additional rectal swab specimens reacted with Coxsackie B antisera but could not be conclusively serotyped. Tissue culture fluids and rectal swab specimens containing other viruses such as ECHO virus, Coxsackie virus A, rhinovirus, rotavirus and Norwalk virus were consistently negative in the assay. The Coxsackie B ELISA offers potential for the rapid identification of Coxsackie B antigens in clinical specimens and tissue culture systems.     

Reverse radioimmunoassays of IgM and IgG antibodies to Coxsackie B viruses in patients with acute myopericarditis

A reverse radioimmunoassay (RIA) of antibodies to enteroviruses, previously developed for the detection of IgM antibodies to Coxsackie B1 (CB1) and B3 (CB3) and to Echo 11 (E11) and 30 (E30) viruses, was extended in the present study for the detection of IgM antibodies to Coxsackie B2 (CB2), B4 (CB4), and B5 (CB5) viruses and of IgG antibodies to CB1-CB5, E11, and E30 viruses. After standardisation of the assays and application to a collection of serum specimens from patients with proven enterovirus infections, specimens from patients with diagnosed or suspected acute myo- and/or pericarditis (myopericarditis group), and control specimens from patients with nonenterovirus infections, were studied, as well as from apparently healthy subjects. Of the patients with enterovirus infections, 29 of 30 (97%) were positive in the IgM RIA and 19 of 25 (76%) in the IgG RIA. In the myopericarditis group, 18 of 37 (49%) patients showed Coxsackie B (CB) virus-specific IgM titres and 9 of 37 (24%) CB virus-specific IgG titres. In the control specimens very few positive responses were detected. The RIAs appeared to be type specific or at least predominantly type specific, provided that the amount of labeled virus was carefully standardised. The sensitivity of the RIAs seemed to be rather high for IgM but low for IgG. In the neutralisation (NT) test no significant rise or fall in titre against CB viruses was demonstrated in the myopericarditis group. It is concluded that the reverse IgM RIA may be valuable for studies of the role of CB viruses in acute myo- and/or pericarditis.     

Immunoglobulin responses to echovirus type 11 by enzyme linked immunosorbent assay: Single-serum diagnosis of acute infection by specific IgM antibody

An indirect solid-phase enzyme-linked immunosorbent assay (ELISA) for the determination of specific IgM and IgG antibodies to echovirus type 11 in a single dilution of serum was developed using partially purified echovirus type 11 bound to microplates. Whole serum was used for IgG antibody but prior to assaying for IgM antibody interfering IgG was removed by ion exchange chromatography. The ELISA for echovirus type 11 IgG antibody was a more sensitive, rapid, technically easier and less costly alternative to the neutralisation test. With the IgG ELISA 12 of 132 sera (10.6%) known to contain enterovirus antibodies other than echovirus type 11 were positive but it could not be determined to what extent this was due to the greater sensitivity of the ELISA or cross-reactions. The IgM ELISA was even more sensitive than the IgG ELISA with acute sera, and showed a reactivity in 4 of 36 sera (11.1%) with no detectable echovirus type 11 neutralising antibodies. Echovirus type 11 IgM antibody was detected in all sera collected after the first week of infection and up to 30 days after infection. However, it was only detected in 58% of sera collected during the first week after onset thus limiting its use for rapid diagnosis. The echovirus type 11 IgM ELISA appears to have considerable laboratory diagnostic potential when a rising antibody level cannot be demonstrated in paired sera or when virus is not cultured.     

Detection of anti-delta antibodies among acute hepatitis B virus-infected patients

A total of 130 acute HBsAg-positive subjects were tested for the presence of delta hepatitis antibodies (anti-HD). Of the 130 subjects, 56 were females and 74 were males. All patients were individuals attending the two teaching hospitals, i.e., Parirenyatwa and Harare Central hospitals. All the sera were examined by the enzyme-linked immunosorbent assay (ELISA), and results were read spectrophotometrically at a wavelength of 492 nm. Of 130, 11 (19.64%) males had anti-HD present in the serum with an overall prevalence of 21 (16.15%). The positivity rates of HBeAg and anti-HBc were 64 (49.23%) and 116 (89.23%), respectively. There was no significant difference between the positive rate in males as compared to females.     

Coxsackie B virus infection in coronary care unit patients

It has been suggested that Coxsackie B virus infections may play a part in causing or triggering myocardial infarction. This study was designed to compare the incidence of such infections in Coronary Care Unit patients and normal controls. The choice of a suitable criterion for diagnosis of Coxsackie infection is discussed fully. Two hundred and fifty admissions to a Coronary Care Unit and 100 control subjects had a serum sample tested by microneutralisation for Coxsackie B antibodies. The incidence of infection among 130 patients diagnosed as acute myocardial infarction was 5% compared with 4% in the control group. In a subgroup classified as non-transmural myocardial infarction, the incidence of infection was 14%. The sex ratio of this group differed from the myocardial infarction group as a whole suggesting that the non-transmural group may not have been homogeneous. Normal coronary arteriograms were subsequently found in three patients who were diagnosed as non-transmural myocardial infarction but who had serological evidence of recent Coxsackie infection. This study does not demonstrate an association between Coxsackie infection and myocardial infarction as a whole and does not support the view that Coxsackie infection causes or provokes myocardial infarction. It does, however, suggest that myocarditis may simulate non-transmural infarction.     

Serum IgA, IgG, and IgM responses to different enteroviruses as measured by a coxsackie B5-based indirect ELISA.

An enterovirus-specific indirect ELISA, based on a single local isolate of coxsackie B5 as antigen, was used to study the IgA, IgG, and IgM responses in 19 patients with a recent or current enterovirus infection. Twelve different enterovirus serotypes were isolated from 15 patients. Paired serum samples were available from 10 and a single serum from 5 of these 15 patients. In addition, 4 patients diagnosed by a significant titer rise of complement fixing antibodies to enterovirus were included. A serological diagnosis, defined as an increase in titer of enterovirus IgG and/or presence of enterovirus IgM, were established in all 14 patients with paired sera. Enterovirus IgM was present in either a single serum or in both sera in 13 of them. Out of 5 patients with a single serum sample only, enterovirus IgA or enterovirus IgM was found in 4. Specific IgA was present in either a single serum or in both sera in 14 of the 19 patients. Seven of the 10 enterovirus isolate patients with paired sera had a significant titer rise of complement fixing antibodies; however, all 10 were diagnosed by ELISA. Among 64 healthy controls 2 had enterovirus IgA and none had enterovirus IgM. In conclusion, the use of a single antigen-based ELISA was found to be reliable for the diagnosis of recent and current enterovirus infections.     

庚型肝炎病毒IgM抗体检测方法的建立及其临床意义

目的建立一种早期、快速诊断庚型肝炎的血清学检测方法。方法以辣根过氧化物酶标记庚型肝炎病毒（ＨＧＶ）多肽ＮＳ３,ＮＳ５区段抗原,建立了捕获酶联免疫吸附试验（ＥＬＩＳＡ）,用于检测血清中ＨＧＶＩｇＭ抗体。结果本法不受特异性ＩｇＧ的竞争和类风湿因子的干扰;与其它致肝炎的病毒（ＨＡＶ、ＨＢＶ、ＨＣＶ、ＨＥＶ、ＣＭＶ、ＥＢＶ）无交叉反应。检测４６例非甲、乙、丙、戊型肝炎患者血清,抗－ＨＧＶＩｇＭ阳性１４例,阳性率３０．４３％,其中,６例同时为ＨＧＶＲＮＡ阳性,阳性符合率为４２．８６％（６／１４）,检测１２例庚型肝炎病人双份血清,其中,急性期血ＨＧＶＩｇＭ抗体均为阳性。结论该法检测ＨＧＶＩｇＭ抗体特异性强,敏感性高,且简便快速,适用于临床对庚型肝炎新近感染的早期诊断,有推广应用价值。     

Measles virus-specific IgM antibodies in sera from patients with chronic active hepatitis

The occurrence of measles virus-specific IgM antibodies in sera from patients with chronic active hepatitis not caused by Hepatitis B virus was examined by a specific enzyme-linked immunosorbent assay (ELISA). Using whole serum, specific IgM antibodies were detected in 12 of 23 sera from patients with hepatitis B surface antigen (HBsAg)-negative chronic active hepatitis. In nine of these sera the finding of specific IgM antibodies was verified by separation of IgM by density gradient centrifugation and examination of the fractions by ELISA. Most of the sera from the patients with measles virus-specific IgM antibodies had an elevated level of specific IgG antibodies compared to the level of IgG found in control sera. The significance of these findings in view of a possible persistent measles virus antigen production in patients with chronic active hepatitis is discussed.     

Detection of IgM antibodies against cytomegalovirus: comparison of two radioimmunoassays, enzyme-linked immunosorbent assay and immunofluorescent antibody test

The sensitivity and specificity of direct antibody radioimmunoassay (RIA), M-antibody capture RIA (MACRIA), enzyme-linked immunosorbent assay (ELISA), and the immunofluorescent antibody (IFA) test for the detection of CMV-specific IgM was compared using 40 sera selected from different groups of patients. RIA, MACRIA, and ELISA gave concordant results with thirty-two sera but discordant results with eight sera, of which three were cord sera from congenitally infected babies, three were from immunocompromised patients with recurrent CMV infections, and two were from patients with lymphadenopathy and Paul-Bunnell-positive mononucleosis, respectively. RIA, MACRIA, and ELISA were of similar sensitivity with sera from adult patients, but ELISA was apparently less sensitive than RIA and MACRIA for the detection of CMV IgM in cord serum. By comparison IFA was significantly less sensitive than the other three tests. Rheumatoid factor is reactive in RIA, ELISA, and IFA but can efficiently be removed by absorption with latex-IgG beads or cross-linked human IgG.     

柯萨奇B3病毒引起心肌细胞凋亡的初步探索

目的 以病毒性心肌炎（ｖｉｒａｌ ｍｙｏｃａｒｄｉｔｉｓ,ＶＭＣ）细胞感染模型为基础,研究柯萨奇Ｂ３病毒（ＣｏｘｓａｃｋｉｅＢ３ ｖｉｒｕｓ,ＣＶＢ３）感染与心肌细胞凋亡的关系。方法 在嗜鼠心肌柯萨奇Ｂ３病毒（ＣＶＢ３ｍ）感染心肌细胞后２４ｈ电镜观察细胞超微结构的改变,同时取细胞悬液以流式细胞仪检测“凋亡峰”,了解细胞亡率,并采用酶联免疫分析法（ＥＬＩＳＡ）定量测定ＣＶＢ３ｍ病毒感染后１、２、４、     

Mu-antibody capture ELISA for the rapid diagnosis of enterovirus infections in patients with aseptic meningitis

Between August and November 1985, 45 patients with suspected aseptic meningitis were investigated using conventional virus isolation procedures and the mu-antibody capture Coxsackie B IgM enzyme-linked immunoabsorbent assay (ELISA) test, which is well known to cross-react with other members of the enterovirus group. An enterovirus was isolated from 22% of patients compared with 67% who were positive in the ELISA test. Not only was the rate of enterovirus detection increased by using this ELISA method, the clinician received a result within 2 days of submission of serum to the laboratory. A positive result was reassuring to the patient and helpful in clinical management. The main disadvantage of this test was its cost since Coxsackie B1-5 virus antigens were essential. Development of a single inexpensive enterovirus-specific antigen is thus desirable.     

Autoantibodies to survivin in patients with chronic hepatitis and hepatocellular carcinoma

Objectives. Autoantibodies to tumor-associated antigens including survivin have been detected in sera from patients with hepatocellular carcinoma (HCC). However, little is known about autoantibody responses to tumor-associated antigens in patients with chronic hepatitis, which strongly predisposes to development of HCC.

Methods. We subjected sera from 57 patients with chronic hepatitis and 29 patients with HCC to an enzyme-linked immunosorbent assay (ELISA) using a full-length recombinant survivin protein. A cutoff value for positivity was determined as the mean absorbance +2SD for sera from healthy volunteers.

Results. In patients with chronic viral hepatitis, elevated anti-survivin antibodies were detected in 10 of 57 sera (17.5%); in HCC patients, such elevation were detected in 7 of 29 sera (24.1%). The levels of anti-survivin antibodies in HCC patients with HCV infection were significantly higher than those in the healthy control and HCC patients with HBV infection. However, there were no significant differences in the levels of anti-survivin antibodies between HCV and HCC patients with HCV infection.

Conclusions. We demonstrated that elevated anti-survivin antibodies were detected for the first time in patients with chronic viral hepatitis. The results suggest that the levels of anti-survivin antibodies have no association with the progression of HCV or HBV to HCC.     

The IgM antibody responses to the core antigen of hepatitis B virus

Little is known about the immunoglobulin class of antibodies to HBcAg. In the present study sera containing anti-HBc were fractionated by sucrose density-gradient centrifugation, and all serum fractions were tested against HBcAg by immunoelectro-osmophoresis. In addition selected fractions were examined by complement fixation test, immune adherence hemagglutination and immune electron microscopy. Anti-HBc activity in IgG serum fractions was demonstrated by all four techniques used, but HBcAg-specific IgM was detected only by immunoelectro-osmophoresis and by immune electron microscopy. In acute hepatitis B, HBcAg-specific IgM was detected for up to eight weeks after the onset of jaundice. It was also found transiently in two patients who developed chronic hepatitis B without an icteric episode and in one out of thirteen patients with HBsAg-positive chronic liver disease, but in none of eight healthy HBsAg carriers. The results suggested that HBc Agspecific IgM is formed transiently in response to primary HBV infection but is generally undetectable in established HBsAg carriers.     

Solid-phase radioimmunoassay determination of virus-specific IgM antibody levels in a follow-up of patients with naturally acquired measles infections

The question of the exact disappearance time or possible persistence of measles-specific IgM antibodies after naturally acquired measles virus infections was studied with a sensitive solid-phase radioimmunoassay (RIA) method. A total of 30 patients were analyzed with follow-up times varying from 4.5 to 8 months; all were measles IgM positive in the first serum specimen obtained after the onset of rash. In 29 of 30 patients, the measles IgM declined to undetectable levels by approximately 90 days. The remaining patient developed postmeasles encephalitis, however, and was found to have a prolonged measles IgM antibody response. For comparison, the measles-specific IgG response was also studied and was found to develop only slightly later than the IgM response, with levels then remaining high and stable up to 8 months later. Although apparent measles IgM antibodies were found in 1 of 64 nonmatched adult controls, they were due to the presence of high levels of IgM-class rheumatoid factor. The data presented indicate that measles IgM antibodies begin to decline soon after the onset of rash and reach negative levels 1 to 3 months later; in complicated infections, however, measles IgM antibody synthesis may not terminate normally.     

Recent changes in prevalence pattern of Japanese encephalitis virus in Hyogo Prefecture, Japan, using a two-dimensional distribution of IgG and IgM class antibody levels in swine sera

Surveillance of antibody to Japanese encephalitis (JE) virus in swine sera was performed during the epidemic season from July to September, 1978 to 1983, in Hyogo Prefecture, Japan, using enzyme-linked immunosorbent assay (ELISA). Two-dimensional distribution analyses of ELISA values for IgG and IgM class antibodies indicated recent changes in the prevalence pattern. Time courses of antibody levels observed from 1978 to 1980 were similar to those in the 1960s of major epidemics, indicating a synchronous infection during the second wave of infection. In 1982 and 1983, however, the JE prevalence pattern implies an incessant infection cycle between vector mosquitoes and pigs during the latter half of the epidemic season. The delay of the occurrence of JE in patients confirms the longer period of exposure to infectious vectors.     

Diagnostic significance of anti-HBc IgM (RIA) in healthy HBsAg carriers and in chronic hepatitis B

The diagnostic significance of IgM antibody against hepatitis B core antigen (anti-HBc) in healthy hepatitis B surface antigen (HBsAg) carriers and in subjects affected by chronic hepatitis B was evaluated. IgM anti-HBc was sought and found in all nine patients examined who were affected by acute HBsAg-positive hepatitis. It was also detected in 2 out of 18 patients with HBsAg-positive chronic persistent hepatitis and in 12 out of 42 patients affected by HBsAg-positive chronic active hepatitis. The absence of this marker was noted in all 26 HBsAg healthy carriers and in the subjects with HBsAg-positive cirrhosis. No relationship was found between the presence of IgM anti-HBc and the degree of inflammatory activity in the patients with HBsAg-positive chronic active hepatitis. A correlation was not found between the presence of IgM anti-HBc and the presence of hepatitis B e antigen (HBeAg) in the same patients. These data show that the absence of IgM anti-HBc may be useful in identifying healthy carriers of HBsAg. The presence of this antibody may be a suitable indication of acute HBsAg-positive hepatitis. In patients with chronic active hepatitis B the presence of IgM anti-HBc cannot be used as diagnostic tool in predicting the severity of liver disease.     

前S1抗原与抗-HBc IgM检测结果探讨

检测患者乙型肝炎病毒前S1抗原（Pre-S1Ag）与抗-HBc IgM的水平,探讨两者与HBV急性感染的相关性,为乙型肝炎的早期诊治提供必要的理论依据。对90份临床标本分别采用PCR技术检测HBV-DNA载量阳性标本,采用固相R IA（SPR IA）检测HBeAg、抗-HBc IgM,采用胶体金免疫层析法检测Pre-S1Ag,并对结果进行分析。结果表明90份HBV-DNAPCR阳性标本中Pre-S1Ag阳性71份,HBeAg阳性60份,抗-HBc IgM阳性14份,其检测结果有显著性差异（P〈0.05）。血清Pre-S1Ag、抗-HBc IgM及HBeAg与HBV复制紧密相关。Pre-S1Ag明显优于抗-HBc IgM及HBeAg。因此,Pre-S1Ag的检测对乙型肝炎临床早期诊断、抗病毒治疗方案的选择和预后判断具有重要意义。     

Responses of IgM for enterovirus 71 infection

A rapid serological assay was developed for detection of specific IgM to enterovirus 71, a human picornavirus that is usually associated with severe central nervous system complications. The sensitivity and specificity of this "in-house" mu-capture enzyme linked-immunosorbent assay was assessed by testing 213 serum samples. With the conventional virus culture as a standard method, the sensitivity and specificity were 91.5 and 93.1%, respectively, for this newly developed immunoassay. This method allows for detection of the IgM responses from the patients either infected by genotype B or genotype C of enterovirus 71. IgM can be detected as early as the second day from the onset of disease. IgM responses exhibit 100% positive rate from enterovirus 71-infected patients with complications, including encephalitis, meningitis, polio-like syndrome, pulmonary edema, and fatal cases. These findings suggest that detection of specific IgM by the use of enzyme linked-immunosorbent assay is a rapid and valuable way for the diagnosis of enterovirus 71 infection.