RNA extraction for quantitative enterovirus RT-PCR: comparison of three methods.

Abstract:

:Quantification of virus-like RNA sequences in biological fluids, like serum and cerebrospinal fluid, requires an RNA extraction method that is both reproducible and fast. Three RNA extraction methods were tested on enteroviruses: (1) the acid guanidine thiocyanate-phenol/chloroform (AGPC) method; (2) a method based on differential precipitation of the RNA and (3) a 'bind-wash-elute' system based on silica-gel membrane binding. The latter two methods yielded a comparable detection limit as measured by RT-PCR ELISA. The detection limit for the AGPC method was 10 times higher. The relative standard deviation for the bind-wash-elute method (3%) was superior to that of the other methods tested (both 20%) and provides a reliable and fast method to extract (viral) RNA from biological fluids for quantification by RT-PCR.

journal_name

J Pharm Biomed Anal

authors

Verheyden B,Thielemans A,Rombaut B,Kronenberger P

doi

10.1016/s0731-7085(03)00312-1

subject

Has Abstract

pub_date

2003-11-24 00:00:00

pages

819-23

issue

4

eissn

0731-7085

issn

1873-264X

pii

S0731708503003121

journal_volume

33

pub_type

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