Quantification of human H1 histamine receptor mRNA from peripheral blood

Study Objective. To develop a reverse transcription (RT)-polymerase chain r eaction (PCR) technique to detect and quantify human histamine(1) (H-1) rec eptor mRNA in peripheral blood. Methods. Primer pairs were based on the human H-1 receptor nucleotide seque nce. A competitive. reference standard (CRS) was developed that used the sa me primers as wild-type mRNA but contained a 92-bp deletion. RT-PCR was per formed with 5 mu g of total RNA obtained from venous blood of six subjects that was added to known concentrations of CRS RNA. Linear regression compar ing wild-type with CRS product was used to quantify wild-type mRNA. Measurements and Main Results. Three subjects had detectable H-1 mRNA, with a range of 31-435 pg. In three subjects PCR product was not detected, alth ough the presence of RNA was confirmed. Redesigned primer pairs showed mRNA to H-1 receptor in two of the remaining subjects, but it was undetectable in the third. Conclusion. RT-PCR can be used to detect and quantify human H1 receptor mRN A from peripheral blood.