Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus
Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus
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DOI：
10.1128/jcm.40.2.372-375.2002
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发表时间：
2002-02-01
影响因子：
9.4
通讯作者：
Morikawa, S
中科院分区：
文献类型：
--
作者：
Saijo, M;Qing, T;Morikawa, S
用含有克里米亚-刚果出血热病毒（CCHFV）核蛋白（rNP）cDNA的表达载体（pKS 336-CCHFV-NP）转染HeLa细胞，建立了连续表达CCHFV核蛋白的细胞系。将这些细胞用作间接免疫荧光（IF）的抗原，以检测CCHFV的免疫球蛋白G抗体。通过使用血清样品检查该IF技术的灵敏度和特异性，并将其与使用CCHFV感染的Vero E6细胞（真实抗原）的IF技术进行比较。在HeLa细胞中表达的CCHFV rNP的染色显示出与CCHFV感染的Vero E6细胞相似的独特颗粒图案。阳性染色可以很容易地与阴性结果区分开。所有13份通过使用真实抗原确定为阳性的血清样品也通过使用表达CCHFV rNP的HeLa细胞（重组抗原）确定为阳性。108份用真实抗原检测为阴性的血清样品，用重组抗原检测也为阴性。因此，该IF技术的灵敏度和特异性与具有真实抗原的IF相比均为100%。使用表达CCHFV rNP的HeLa细胞的新型IF技术不仅可用于CCHF的诊断，还可用于CCHFV感染的流行病学研究。
A HeLa cell line continuously expressing recombinant nucleoprotein (rNP) of the Crimean-Congo hemorrhagic fever virus (CCHFV) was established by transfection with an expression vector containing the cDNA of CCHFV NP (pKS336-CCHFV-NP). These cells were used as antigens for indirect immunofluorescence (IF) to detect immunoglobulin G antibodies to CCHFV. The sensitivity and specificity of this IF technique were examined by using serum samples and were compared to those of the IF technique using CCHFV-infected Vero E6 cells (authentic antigen). Staining of the CCHFV rNP expressed in HeLa cells showed a unique granular pattern similar to that of CCHFV-infected Vero E6 cells. Positive staining could easily be distinguished from a negative result. All 13 serum samples determined to be positive by using the authentic antigen were also determined to be positive by using CCHFV rNP-expressing HeLa cells (recombinant antigen). The 108 serum samples determined to be negative by using the authentic antigen were also determined to be negative by using the recombinant antigen. Thus, both the sensitivity and the specificity of this IF technique were 100% compared to the IF with authentic antigen. The novel IF technique using CCHFV rNP-expressing HeLa cells can be used not only for diagnosis of CCHF but also for epidemiological studies on CCHFV infections.