Real-time nucleic acid sequence-based amplification using molecular beacons for detection of enterovirus RNA in clinical specimens.
Real-time nucleic acid sequence-based amplification (NASBA) using molecular beacon technology (NASBA-beacon) was compared to standard NASBA with postamplification hybridization using electrochemiluminescently labeled probes (NASBA-ECL) for detection of enteroviruses (EV) in 133 cerebrospinal fluid and 27 stool samples. NASBA-ECL and NASBA-beacon were similar in sensitivity, detecting 55 (100%) and 52 (94.5%) EV-positive samples, respectively. There were no false positives. Both NASBA assays were significantly more sensitive than culture. Real-time NASBA-beacon reagents and equipment rental were more expensive than those for NASBA-ECL; however, time to result was shortened by 1.5 h, hands-on time was reduced by 25 min, and the assay was much simpler for technologists to learn and perform.
Landry
Marie L
ML
Department of Laboratory Medicine, P.O. Box 208035, Yale University School of Medicine, New Haven, CT 06520-8035, USA. marie.landry@yale.edu
Garner
Robin
R
Ferguson
David
D
D000293:Adolescent; D000328:Adult; D002555:Cerebrospinal Fluid / Q000821:virology; D002648:Child; D004563:Electrochemistry / Q000379:methods; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000821:virology*; D005243:Feces / Q000821:virology; D006801:Humans; D007223:Infant; D008163:Luminescent Measurements / Q000379:methods; D008875:Middle Aged; D015335:Molecular Probes; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity
D015335:Molecular Probes; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic