Attoui H, Billoir F, Bruey J M, de Micco P, de Lamballerie X
Laboratoire de Virologie Moléculaire, Tropicale et Transfusionnele, Faculté de Médecine de Marseille, France.
Am J Trop Med Hyg. 1998 Nov;59(5):763-8. doi: 10.4269/ajtmh.1998.59.763.
Molecular and serologic methods usable for the biological diagnosis of Coltivirus infection are reported. We designed a multiplex reverse transcription-polymerase chain reaction system that allowed the simultaneous and specific amplification of three genomic segments from as little as 0.01 plaque-forming units. Another system in the S2 viral segment permitted the differential diagnosis of American and European viral isolates. We also discuss some improvements of previous ELISAs, and the results obtained with paired sera from Colorado tick fever (CTF) virus-infected individuals. Western blot analysis was developed that allowed the detection of antibodies to a 38-kD viral protein in all tested sera. It also enabled the detection of anti-CTF virus antibodies in ELISA-negative sera. Specific IgM antibodies against a synthetic viral peptide could be detected in sera at the acute stage of the infection. Together, these results should permit the diagnosis of Coltivirus infection at any stage of the pathology.
报告了可用于科蒂病毒感染生物学诊断的分子和血清学方法。我们设计了一种多重逆转录-聚合酶链反应系统，该系统能够从低至0.01个噬斑形成单位同时特异性扩增三个基因组片段。S2病毒片段中的另一个系统可用于鉴别诊断美洲和欧洲的病毒分离株。我们还讨论了先前酶联免疫吸附测定（ELISA）的一些改进，以及从感染科罗拉多蜱传热（CTF）病毒的个体获得的配对血清的检测结果。开发了蛋白质印迹分析方法，可在所有测试血清中检测到针对一种38-kD病毒蛋白的抗体。它还能够在ELISA阴性血清中检测到抗CTF病毒抗体。在感染急性期的血清中可检测到针对合成病毒肽的特异性IgM抗体。这些结果共同应能在病理学的任何阶段诊断科蒂病毒感染。
