[Riboprobes: an alternative in the detection of viral sequences].
The use of riboprobes for the detection of RNA viral sequences is analyzed. Subgenomic fragments of cDNA from poliovirus type 2, dengue virus type 4 and human immunodeficiency virus type 1, were inserted downstream SP6 and/or T7 promoters in the transcription vectors pGEM-4z or pSP64. RNAs obtained by in vitro transcription in the presence of UTP infinity (32P) were used as probes for the detection of RNA viral sequences from infected cell lines in slot and Northern blot assays. The poliovirus riboprobe (P2-221) was able to detect specific viral sequences; thus, it could be used for the detection of the virus in serum, as well as in residual waters. The human immunodeficiency virus riboprobe (HIV-378), detected viral sequences poly A+RNA from infected cells; thus it can be used as a confirmatory test or as a tool in basic research. Finally, the dengue virus riboprobes (D4-2819 and D4-1134) detected specifically dengue 4 virus; however the sensitivity of the detection could be significantly improved amplifying viral sequences by the polymerase chain reaction (PCR) prior to probe hybridization.
Gutiérrez-Escolano
A L
AL
Laboratorio del Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados, Instituto Politécnico Nacional (CINVESTAV-IPN), México.
Castro-Rodríguez
E M
EM
Núñez-de Cáceres
R M
RM
Soler-Claudín
C
C
Fernández-Tomás
C
C
D015152:Blotting, Northern; D002478:Cells, Cultured; D003715:Dengue / Q000175:diagnosis; D003716:Dengue Virus / Q000235:genetics*; D015658:HIV Infections / Q000175:diagnosis; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000235:genetics; D011051:Poliomyelitis / Q000175:diagnosis; D017955:Poliovirus / Q000235:genetics*; D015347:RNA Probes / Q000096:biosynthesis; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D014776:Virus Cultivation
D015347:RNA Probes; D012367:RNA, Viral
Las ribosondas: una alternativa en la detección de secuencias virales.