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A highly sensitive chemiluminescent reverse transcriptase assay for human immunodeficiency virus. 2002
Fumitomo Odawara, and
Hideki Abe, and
Takehiro Kohno, and
Youko Nagai-Fujii, and
Kenji Arai, and
Shigeyuki Imamura, and
Hideo Misaki, and
Hiroshi Azuma, and
Kenji Ikebuchi, and
Hisami Ikeda, and
Sharad Mohan, and
Kouichi Sano
Fine Chemicals and Diagnostic Division, Asahi Kasei Corporation, 632-1 Mifuku, Ohito-cho, Tagata-gun, Shizuoka 410-2321, Japan. odawara.fb@om.asahi-kasei.co.jp
A simple and highly sensitive reverse transcriptase (RT) assay was developed by combining a previously reported non-radioisotopic RT assay with the use of a template-primer-immobilized microplate, an enzyme capture protocol, product digestion and a chemiluminescent substrate. The assay was able to detect directly the RT activity in serum samples, plasma and cell culture medium without the need for concentration and extraction of the enzyme. The assay was able to detect RT activity equivalent to 100 virions/ml of HIV-1. These results suggest that this highly sensitive chemiluminescent RT assay can be used not only for virological investigation but also for routine screening of biopharmaceuticals.
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