A QUANTITATIVE POLYMERASE CHAIN-REACTION ASSAY FOR INTERLEUKIN-5 MESSENGER-RNA

被引：17

作者：

GUIFFRE, A

ATKINSON, K

KEARNEY, P

机构：

[1] Department of Hematology, St. Vincent's Hospital, Rushcutter's Bay

来源：

ANALYTICAL BIOCHEMISTRY | 1993年 / 212卷 / 01期

关键词：

D O I：

10.1006/abio.1993.1290

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The quantitation of low abundance messenger (m)RNA species such as that for interleukin 5 (IL-5) cannot be achieved by Northern blotting. Polymerase chain reaction (PCR) technology can be used to overcome this paucity of target molecules, but in its standard format it is semiquantitative. We describe the synthesis, purification, and use of a synthetic IL-5 mRNA (cRNA) molecule which, when included in the PCR reaction, enabled the amplification of IL-S mRNA molecules from clinical samples to be made quantitative. A stringent protocol involving analysis of the purified transcript by PCR (to ensure its freedom from template molecules) is required. Preliminary experiments indicate that IL-S mRNA levels are elevated in asthmatics. © 1993 Academic Press, Inc.

引用

页码：50 / 57

页数：8

共 25 条

[1]

ATKINSON K, 1992, BONE MARROW TRANSPL, V9, P175

[2]

BACCHETTA R, 1990, J IMMUNOL, V144, P902

[3] ABSOLUTE MESSENGER-RNA QUANTIFICATION USING THE POLYMERASE CHAIN-REACTION (PCR) - A NOVEL-APPROACH BY A PCR AIDED TRANSCRIPT TITRATION ASSAY (PATTY) [J].