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РЖ ВИНИТИ 34 (BI39) 95.12-04К1.12

Quantitation of IL-1'бета' mRNA by a combined method of RT-PCR and an ELISA based on ion-sensitive field effect transistor [Text] / Hiroshi Tsuruta [et al.] // J. Immunol. Meth. - 1995. - Vol. 180, N 2. - P259-264 . - ISSN 0022-1759
Перевод заглавия: Количественное определение мРНК интерлейкина-1'бета' при сочетании методов РТ-реакции полимеразной цепи и твердофазного иммуноферментного основано на транзиторном влиянии ион-чувствительного поля
Аннотация: A method for quantitative RT-PCR using ELISA detection was developed and applied to the quantitation of IL-1'бета' mRNA in clinical samples. To compensate for the `tube effect' of RT-PCR, asynthetic RNA, pRSET RNA, was added to sample solutions as an internal standard and co-amplified with IL-1'бета' mRNA. Sense primers for IL-1'бета' and pRSET were labeled with digoxigenin and FITC, respectively, while anti-sense primers for both were labeled with biotin. Double-stranded PCR products were captured by two solid-phase pipettetips. One was coated with an anti-digoxigenin antibody and another with an anti-FITC antibody, and sandwiched by avidin-urease, and their activities were measured by coupling them with a pH-FET in a pH-measuring cell containing urea solution. The ratio of the signal intensity for IL-1'бета' to that for pRSET was used to quantify the concentration of IL-1'бета' mRNA. A calibration curve was obtained by using a known amount of AW 109 RNA as an external standard of IL-1'бета'RNA. It was found that 10{2}-10{6} copies of IL-1'бета' mRNA were measurable by the present method. Expression levels of IL-1'бета' mRNA in clinical samples, such as monocytes of peripheral blood or synovial cells from patients with RA or OA, were determined. Япония, Centr. Res. Labs., Kurachiki. Библ. 11

ГРНТИ	34.43.05

ВИНИТИ 341.43.05.07
Рубрики: ИНТЕРЛЕЙКИН 1 БЕТА
КОНТРОЛИРУЮЩАЯ МРНК
СОЧЕТАННЫЙ МЕТОД КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ
РЕАКЦИЯ ПОЛИМЕРАЗНОЙ ЦЕПИ
ТВЕРДОФАЗНЫЙ ИММУНОФЕРМЕНТНЫЙ МЕТОД
ЧЕЛОВЕК

Доп.точки доступа:
Tsuruta, Hiroshi; Matsui, Satomi; Oka, Keiko; Namba, Toshihiko; Shinngu, Masao; Nakamura, Michihiro

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