D. Bresters et al., DURATION OF FIXATION INFLUENCES THE YIELD OF HCV CDNA-PCR PRODUCTS FROM FORMALIN-FIXED, PARAFFIN-EMBEDDED LIVER-TISSUE, Journal of virological methods, 48(2-3), 1994, pp. 267-272
The extent of hepatitis C virus (HCV) RNA loss during increasing perio
ds of fixation of liver tissue in formalin was examined. For this purp
ose human liver tissue, known to be HCV RNA positive and stored at -70
degrees C until use, was cut into small slices (n = 9), which were fi
xed in phosphate-buffered formalin for increasing periods of time befo
re embedding in paraffin. Nucleic acids were extracted from each slice
of formalin-fixed, paraffin-embedded liver tissue and HCV RNA loss du
ring fixation was semi-quantified by testing 10-fold dilutions of each
extract in an HCV cDNA-PCR assay. The endpoint dilution for HCV RNA d
etection by cDNA-PCR in liver slices fixed in buffered formalin for 8-
24 h was comparable to the endpoint dilutions found for 'fresh', non-f
ixed liver slices. After fixation for 2-3 days the endpoint dilution f
or HCV RNA detection was 10(2) to 10(3)-fold less. After 2-4 weeks of
formalin-fixation, HCV RNA was detectable from undiluted nucleic acid
extracts only. It is concluded that formalin-fixed, paraffin-embedded
liver biopsies can be used for HCV RNA detection by cDNA-PCR, on condi
tion that the liver tissue has been embedded in paraffin within 24 h o
f formalin-fixation.