Detection of poly(I):poly(C12U), mismatched double-stranded RNA, by rapid solution hybridization: blood values after intravenous infusion.
A rapid solution hybridization technique has been developed for estimating blood concentrations of poly(I):poly(C12U), a high molecular weight bioactive double-stranded RNA. Samples were prepared by mixing 100 microL of blood with, 165 microL of 6 M guanidine thiocyanate (GuSCN) and 0.16 M EDTA pH 8.0, and freezing. Hybridizations were carried out with a [3H]poly(C) probe in 3 M GuSCN for 10 min at 37 degrees C. Ribonuclease-resistant hybrids were collected by precipitation with trichloroacetic acid and filtration. Validation studies demonstrated minor interassay variance; the assay was accurate in the range 0.1 to 10 ng poly(I):poly(C12U). Thirty-one blood samples from 15 patients were collected and prepared before and immediately after an average 35 min intravenous infusion of 40-500 mg poly(I):poly(C12U). Postadministration values averaged 48% (s.d. 23%) of the theoretical maximum (range 20-102%). These results confirm previous observations of rapid elimination kinetics of poly(I):poly(C12U) in patients.
Strauss
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Hahnemann University, Department of Neoplastic Diseases, Philadelphia, PA 19102.
Strayer
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DR
Gillespie
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DH
D000328:Adult; D000368:Aged; D005260:Female; D006146:Guanidines; D006801:Humans; D007262:Infusions, Intravenous; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D009369:Neoplasms / Q000097:blood; D009693:Nucleic Acid Hybridization; D011070:Poly I-C / Q000097:blood*; D011072:Poly U / Q000097:blood*; D012330:RNA, Double-Stranded / Q000097:blood*; D012260:Ribonucleases / Q000032:analysis; D013861:Thiocyanates
D006146:Guanidines; D012330:RNA, Double-Stranded; D013861:Thiocyanates; D011072:Poly U; C054436:guanidine thiocyanate; C047490:poly(I).poly(c12,U); D012260:Ribonucleases; D011070:Poly I-C