Descôteaux J P, Lussier G, Berthiaume L, Alain R, Seguin C, Trudel M
Arch Virol. 1985;84(3-4):241-50. doi: 10.1007/BF01378976.
The immunoelectron microscopy (IEM) technique has been used for the detection of a rabbit enteric coronavirus (RECV). Immune serum was prepared in guinea pigs; the viral antigen used for the immunization procedure was obtained from the caecum of a sick rabbit, concentrated by centrifugation and purified on Percoll gradient. In order to identify the viral particles used in the immunization procedure, the protein pattern of the particles was determined by electrophoresis and compared with the pattern of other known coronaviruses. Analysis of structural polypeptides of the purified viral particles revealed a pattern similar to that reported for other coronaviruses. These polypeptides cross reacted with two other coronavirus specific immune sera (IBV and TGE). IEM assay of fecal samples collected from healthy and sick rabbits showed the presence of immune aggregates in specimens from both sick and healthy rabbits. Those aggregates contained viral particles sharing morphological characteristics with other coronaviruses. Furthermore, IEM assay was shown to be more sensitive than a direct EM procedure to detect coronavirus particles in rabbit feces. This assay also allowed the detection of a larger number of chronic carriers.
免疫电子显微镜(IEM)技术已用于检测兔肠道冠状病毒(RECV)。在豚鼠中制备免疫血清;用于免疫程序的病毒抗原取自患病兔子的盲肠,通过离心浓缩并在Percoll梯度上纯化。为了鉴定免疫程序中使用的病毒颗粒,通过电泳确定颗粒的蛋白质图谱,并与其他已知冠状病毒的图谱进行比较。对纯化病毒颗粒的结构多肽分析显示出与其他冠状病毒报道的图谱相似的图谱。这些多肽与另外两种冠状病毒特异性免疫血清(IBV和TGE)发生交叉反应。对从健康和患病兔子收集的粪便样本进行IEM检测,结果显示患病和健康兔子的样本中均存在免疫聚集体。这些聚集体包含与其他冠状病毒具有形态学特征相同的病毒颗粒。此外,IEM检测在检测兔粪便中的冠状病毒颗粒方面比直接电子显微镜检查程序更敏感。该检测还能够检测到更多的慢性携带者。