Nested polymerase chain reaction for high-sensitivity detection of enteroviral RNA in biological samples.
A method based on nested polymerase chain reaction was developed for the detection of enteroviral genomes in biological samples. By taking advantage of the conserved 5' noncoding region of the enteroviral RNA, two sets of primers were utilized, enabling the detection either of a broad range of enteroviruses or of group B coxsackieviruses only. The sensitivity of the method is close to the detection of single molecules of viral RNA in as much as 1 mg of tissue sample. A preliminary study showed the usefulness of this technique for the analysis of endomyocardial biopsy samples from patients with idiopathic dilated cardiomyopathy and myocarditis.
Severini
G M
GM
International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
Mestroni
L
L
Falaschi
A
A
Camerini
F
F
Giacca
M
M
D001483:Base Sequence; D002311:Cardiomyopathy, Dilated / Q000382:microbiology; D004279:DNA, Viral / Q000235:genetics; D004770:Enterovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D029822:Enterovirus B, Human / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D006321:Heart / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D009205:Myocarditis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014773:Virology / Q000379:methods / Q000706:statistics & numerical data
D004279:DNA, Viral; D012367:RNA, Viral