Chen M Y, Lee K L, Hung C C
Department of Internal Medicine, National Taiwan University Hospital, Taipei.
J Formos Med Assoc. 2000 Jan;99(1):24-32.
To identify parvovirus B19 infection by means of immunoglobulin (Ig) G and IgM immunoblots among immunocompetent patients who tested negative or had low-titer B19 IgM antibodies in enzyme-linked immunosorbent assays (ELISA).
Serum samples were obtained from 20 patients with parvovirus B19 infection. Another 130 study subjects presumed to be without B19 infection (40 medical personnel and 90 prisoners) were also included. All sera from the patient and study groups tested positive for IgG or IgM with ELISA and were further evaluated using the immunoblot method. Detection of B19 DNA by nested polymerase chain reaction (PCR) was also performed on IgG and IgM positive sera.
IgM immunoblots disclosed one false positive IgM ELISA result in the patient group and three false positive results in the study group. In the patient group, four patients were in the latter stage of antibody response to B19 infection as suggested by the low titer of anti-B19 IgM, incomplete IgM immunoblots, with only a weak viral capsid protein VP-N reaction band, and fading but still strong reaction bands on IgG immunoblots. Strong reaction bands on IgG immunoblots comparable to these four patients were found in three of the 130 study group sera. Furthermore, B19 DNA was detected in three of the four patients and one of the three study subjects by means of nested-PCR. A serum sample from one study subject showed strong IgG but no IgM reactivity to viral capsid protein VP2; nested PCR identified B19 DNA in this serum sample.
Immunoblots and nested PCR should be applied in the diagnosis of B19 infection for patients with low-titer anti-B19 IgM tested by means of ELISA. For diagnosis of B19 infections in certain clinical entities such as chronic arthritis of recent onset and hydrops fetalis, B19 IgM antibodies may have disappeared but B19 infection can still be recognized by the intensity of the reaction bands on IgG immunoblots. The correlation between chronic B19 infection and persistence of antilinear VP2 epitopes requires further study.
通过免疫球蛋白(Ig)G和IgM免疫印迹法,在酶联免疫吸附测定(ELISA)中检测为阴性或B19 IgM抗体滴度较低的免疫功能正常患者中识别细小病毒B19感染。
采集20例细小病毒B19感染患者的血清样本。另外还纳入了130名推测无B19感染的研究对象(40名医务人员和90名囚犯)。患者组和研究组的所有血清经ELISA检测IgG或IgM均为阳性,并进一步采用免疫印迹法进行评估。对IgG和IgM阳性血清也进行巢式聚合酶链反应(PCR)检测B19 DNA。
IgM免疫印迹显示患者组有1例假阳性IgM ELISA结果,研究组有3例假阳性结果。在患者组中,4例患者处于对B19感染抗体反应的后期,表现为抗B19 IgM滴度低、IgM免疫印迹不完整,仅病毒衣壳蛋白VP - N反应带较弱,而IgG免疫印迹上的反应带逐渐褪色但仍较强。在130名研究组血清中的3例中发现了与这4例患者IgG免疫印迹上类似的强反应带。此外,通过巢式PCR在4例患者中的3例以及3名研究对象中的1例检测到了B19 DNA。1名研究对象的血清样本显示对病毒衣壳蛋白VP2有强IgG反应但无IgM反应;巢式PCR在该血清样本中鉴定出B19 DNA。
对于ELISA检测抗B19 IgM滴度较低的患者,免疫印迹法和巢式PCR应用于B19感染的诊断。对于某些临床病症如近期发作的慢性关节炎和胎儿水肿的B19感染诊断,B19 IgM抗体可能已经消失,但通过IgG免疫印迹上反应带的强度仍可识别B19感染。慢性B19感染与抗线性VP2表位持续存在之间的相关性需要进一步研究。