Chemiluminescent enzyme-linked immunoassay for reverse transcriptase, illustrated by detection of HIV reverse transcriptase.
Chemiluminescent enzyme-linked immunoassay for reverse transcriptase, illustrated by detection of HIV reverse transcriptase.
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通过检测 HIV 逆转录酶来说明逆转录酶的化学发光酶联免疫分析。
DOI：
10.1006/abio.1993.1196
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发表时间：
1993
影响因子：
2.9
通讯作者：
Steigbigel,RT
中科院分区：
文献类型：
--
作者：
Suzuki,K;Craddock,BP;Kano,T;Steigbigel,RT
用生物素标记的寡聚脱氧胸腺酸和地高辛-脱氧尿嘧啶核苷三磷酸代替氚胸苷三磷酸，建立了检测人类免疫缺陷病毒1型逆转录酶的化学发光分析方法。RT反应后，新聚合的生物素寡聚脱氧核糖核酸含有地高辛标记。该核苷酸通过与生物素的反应结合到包被链霉亲和素的微滴定板上。在检测步骤中，加入地高辛配基的碱性磷酸酶偶联抗体，然后与该酶的化学发光底物反应。该方法与用纯化的禽成髓细胞增生症病毒逆转录酶(RT)进行同位素测定具有非常密切的相关性。该方法还与体外感染HTLV-IIIB的淋巴细胞的同位素RT方法进行了比较，再次证明了该方法具有较高的相关性。RT反应后的总检测时间小于100min。
A chemiluminescent assay for reverse transcriptase (RT) of the human immunodeficiency virus 1 was developed using biotin-labeled oligodeoxythymidylic acid (biotin oligo-dT) and digoxigenin-deoxyuridine triphosphate instead of tritiated thymidine triphosphate. After the RT reaction, the newly polymerized strand from biotin oligo-dT contained digoxigenin labels. This nucleotide was bound to a streptavidin-coated microtiter plate by the reaction to biotin. At the detection step, an alkaline phosphatase-conjugated antibody to digoxigenin was added, followed by the reaction of a chemiluminescent substrate for this enzyme. This method shows very close correlation with the isotopic assay using purified avian myeloblastosis virus reverse transcriptase (RT). This assay was also compared with the isotopic RT assay using lymphocytes infected in vitro with HTLV-IIIB and again demonstrated a close correlation. The total assay time after the RT reaction step was less than 100 min.