Development of NASBA ®, a nucleic acid amplification system, for identification of Listeria monocytogenes and comparison to ELISA and a modified FDA method

NASBA ®, an isothermal nucleic acid amplification system was used for identification of Listeria monocytogenes. A primer set and a species-specific probe were selected from the 16S rRNA sequence. The probe was shown to hybridize specifically to the amplifiied single-stranded RNA of L. monocytogenes....

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Bibliographic Details
Published in:International journal of food microbiology 1995-09, Vol.27 (1), p.77-89
Main Authors: Uyttendaele, M., Schukkink, R., van Gemen, B., Debevere, J.
Format: Article
Language:English
Subjects:
ALIMENTOS
Amplification
Bacteriological Techniques
Base Sequence
Biological and medical sciences
ELISA
Enzyme-Linked Immunosorbent Assay
Food industries
Food Microbiology
FOODS
Fundamental and applied biological sciences. Psychology
IDENTIFICACION
IDENTIFICATION
LISTERIA MONOCYTOGENES
Listeria monocytogenes - isolation & purification
Molecular Sequence Data
NASBA
Nucleic Acid Amplification Techniques
Nucleic Acid Hybridization
nucleic acids
PRODUIT ALIMENTAIRE
RNA
RNA, Bacterial
Sensitivity and Specificity
TEST ELISA
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Summary:NASBA ®, an isothermal nucleic acid amplification system was used for identification of Listeria monocytogenes. A primer set and a species-specific probe were selected from the 16S rRNA sequence. The probe was shown to hybridize specifically to the amplifiied single-stranded RNA of L. monocytogenes. No hybridization occured with amplification product of L. seeligeri, L. innocua, L. ivanovii, and L. welshimeri. Detection sensitivity for the NASBA ® assay was determined at 10 6 cfu ml . The possibility of using the NASBA ® assay for detection of L. monocytogenes in foods after a 2-day enrichment procedure was explored. NASBA ® was compared to a modified FDA method and ELISA for detection of L. monocytogenes artificially inoculated (1–100 cfu/25 g) in eight food products. False-negative results were obtained with the modified FDA method (6.75%). NASBA ® and ELISA were shown in this study to detect the pathogen with equal efficiency (no false negative or false positive results). Both methods allowed detection of less than 10 cfu/25 g within 3 days but ELISA can only be used for diagnosis of Listeria spp. while the NASBA ® procedure permitted specific identification of the human pathogen L. monocytogenes.
ISSN:0168-1605
1879-3460
DOI:10.1016/0168-1605(95)00166-H