Use of the colonic carcinoma cell line CaCo-2 for in vivo amplification and detection of enteric viruses.
The use of the continuous cell line CaCo-2 as an in vivo amplification system for the detection of fastidious human enteric viruses is reported. CaCo-2 cells showed an increased sensitivity to laboratory strains of group A rotavirus 3, reovirus 3, astrovirus 1, poliovirus 1, coxsackievirus A 24, enterovirus 70, and adenovirus 5, 40 and 41, when compared to a routine host cell line for each virus. Nucleic acids from wild-type infectious rotavirus, astrovirus, and adenovirus 40 in stool samples of patients with acute gastroenteritis could be amplified after infection of CaCo-2 cells with trypsin-pre-treated virus inocula. Virus diagnosis was carried out subsequently by dot-blot hybridisation with specific cDNA probes. An amplification factor between 10 and 1,000x was obtained by infection of CaCo-2 cells, thus enabling specific detection of low numbers of a wide range of enteric viruses, and the differentiation between infectious and noninfectious particles.
Department of Microbiology, University of Barcelona, Spain.
R M
RM
Pintó
J M
JM
Diez
A
A
Bosch
D000230:Adenocarcinoma; D000256:Adenoviridae / Q000254:growth & development* / Q000302:isolation & purification; D004279:DNA, Viral / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D006801:Humans; D007421:Intestine, Small / Q000821:virology; D001257:Mamastrovirus / Q000254:growth & development* / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification; D012401:Rotavirus / Q000254:growth & development* / Q000302:isolation & purification; D014407:Tumor Cells, Cultured; D014776:Virus Cultivation / Q000379:methods*
D004279:DNA, Viral; D012367:RNA, Viral