Prior Enrichment of HIV-DNA with Probe-DNA Particles for an Efficient PCR Diagnosis
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- FAN Kejun
- Japan Synthetic Rubber Co., Ltd. Tsukuba Resaerch Laboratory
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- TANO Hiroyuki
- Japan Synthetic Rubber Co., Ltd. Tsukuba Resaerch Laboratory
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- KITAJIMA Masaaki
- Japan Synthetic Rubber Co., Ltd. Tsukuba Resaerch Laboratory
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- TAMATSUKURI Shigeru
- Nippon Roche K.K.
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- FURUICHI Yasuhiro
- Nippon Roche K.K.
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- HAYASHI Takako
- AGENE Research Institute
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- IMAI Mitsunobu
- Kanagawa Prefectural Institute of Health
Bibliographic Information
- Other Title
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- プローブDNA結合粒子によるHIV-DNAの特異的な濃縮及びそのPCR検出
- プローブ DNA ケツゴウ リュウシ ニヨル HIV DNA ノ トクイテキ
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Abstract
PCR mediated detection of HIV-DNA has been widely used. However, compared with traditional immunological diagnoses, the extraction of DNA is a laborious and time consuming step. We have developed a procedure for the efficient isolation and concentration of HIV-DNA from cell lysates. We report here a novel method by which one can recover HIV-DNA in a small volume (-50μl) of solution from a large volume of crude cell lysate which contains as few as several copies. The method uses the specific hybridization of HIV-DNA to HIV probe-DNA particles. This prior enrichment augmented the sensitivity in the detection of HIV-DNA by PCR, and allows us to make a diagnosis even if the specimen contained an extremely low copy number of HIV-DNA molecules in a large volume, which would have otherwise resulted in false-negative data with the conventional extraction method. The method also enables the examination of 100 individual blood specimens in a combined form. Thus, the application of the present enrichment procedure with HIV probe-DNA particles should reduce the labor and cost of HIV diagnosis, since the HIV positive samples represent a very minor group of people among specimens subjected to clinical laboratory tests and, particularly, among blood samples voluntarily donated to be used for transfusions.
Journal
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- Kansenshogaku Zasshi
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Kansenshogaku Zasshi 69 (9), 957-962, 1995
The Japanese Association for Infectious Diseases
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Details
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- CRID
- 1390282680022315648
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- NII Article ID
- 10008727072
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- NII Book ID
- AN00047715
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- ISSN
- 1884569X
- 03875911
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- NDL BIB ID
- 3644806
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- PubMed
- 7594795
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed