Use of internal standard RNA molecules for the RT-PCR amplification of the faeces-borne RNA viruses.
The diagnostic system based on reverse transcription (RT)-PCR has been used widely for the detection of viral genomes of faecal-borne RNA viruses. However, faecal specimens often produce both false positive and false negative results. Therefore, there is a need for a diagnosis procedure that can control for 'false-results'. In this study, an internal standard RNA that can serve as a non-competitive positive template was developed and used directly to detect faecal-borne RNA viruses without noticeable competitive inhibition of the target viral genome. These results suggest that the internal standard RNA is a useful standard molecule when undertaking diagnostic qualitative RT-PCR procedures for enteroviruses and related faecal-borne RNA viruses.
Kim
Kisoon
K
Department of Life Science, College of Natural Science, Chung-Ang University, Heuksuk-dong 221, Dongjak-ku, Seoul 156-756, Republic of Korea.
Park
Jeongkoo
J
Chung
Yoonseok
Y
Cheon
Doosung
D
Lee
Il-Beom
IB
Lee
Seokgyu
S
Yoon
Jaedeuk
J
Cho
Haewol
H
Song
Chulyong
C
Lee
Kwang-Ho
KH
D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D005243:Feces / Q000821:virology*; D006801:Humans; D008969:Molecular Sequence Data; D012327:RNA Virus Infections / Q000175:diagnosis*; D012328:RNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification; D012400:Rotavirus Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral