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.
Department of Laboratory Medicine, P.O. Box 208035, Yale University School of Medicine, New Haven, CT 06520-8035, USA. marie.landry@yale.edu
Marie L
ML
Landry
Robin
R
Garner
David
D
Ferguson
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