Development of a Cost-Effective Line Probe Assay for Rapid Detection and Differentiation of
Diagnosis
Line Probe Assay (LPA)
Mycobacterium Infection
Tuberculosis
Journal
Reports of biochemistry & molecular biology
ISSN: 2322-3480
Titre abrégé: Rep Biochem Mol Biol
Pays: Iran
ID NLM: 101637937
Informations de publication
Date de publication:
Jan 2020
Jan 2020
Historique:
entrez:
26
6
2020
pubmed:
26
6
2020
medline:
26
6
2020
Statut:
ppublish
Résumé
The line probe assay (LPA) is one of the most accurate diagnostic tools for detection of different One pair of biotinylated primers and 15 designed DNA oligonucleotide probes were used based on multiple aligned internal transcribed spacer (ITS) sequences. Specific binding of the PCR-amplified products to the probes immobilized on nitrocellulose membrane strips was evaluated by the hybridization method. Experiments were performed three times on separate days to evaluate the assay's repeatability. The PCR-LPA was evaluated directly on nine clinical samples and their cultivated isolates. All 15 probes used in this study hybridized specifically to ITS sequences of the corresponding standard species. Results were reproducible for all the strains on different days. Mycobacterium species of the nine clinical specimens and their cultivated isolates were correctly identified by PCR-LPA and confirmed by sequencing. In this study, we describe a PCR-LPA that is readily applicable in the clinical laboratory. The assay is fast, cost-effective, highly specific, and requires no radioactive materials.
Sections du résumé
BACKGROUND
BACKGROUND
The line probe assay (LPA) is one of the most accurate diagnostic tools for detection of different
METHODS
METHODS
One pair of biotinylated primers and 15 designed DNA oligonucleotide probes were used based on multiple aligned internal transcribed spacer (ITS) sequences. Specific binding of the PCR-amplified products to the probes immobilized on nitrocellulose membrane strips was evaluated by the hybridization method. Experiments were performed three times on separate days to evaluate the assay's repeatability. The PCR-LPA was evaluated directly on nine clinical samples and their cultivated isolates.
RESULTS
RESULTS
All 15 probes used in this study hybridized specifically to ITS sequences of the corresponding standard species. Results were reproducible for all the strains on different days. Mycobacterium species of the nine clinical specimens and their cultivated isolates were correctly identified by PCR-LPA and confirmed by sequencing.
CONCLUSION
CONCLUSIONS
In this study, we describe a PCR-LPA that is readily applicable in the clinical laboratory. The assay is fast, cost-effective, highly specific, and requires no radioactive materials.
Types de publication
Journal Article
Langues
eng
Pagination
383-393Références
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