Evaluation of a commercial probe assay for detection of rifampin resistance inMycobacterium tuberculosis directly from respiratory and nonrespiratory clinical samples

1998 ◽  
Vol 17 (3) ◽  
pp. 189-192
Author(s):  
F. Gamboa ◽  
P. J. Cardona ◽  
J. M. Manterola ◽  
J. Lonca ◽  
L. Matas ◽  
...  
Keyword(s):  
Clinical Samples ◽  
Rifampin Resistance ◽  
Probe Assay

scholarly journals Padlock Probe Assay for Detection and Subtyping of Seasonal Influenza

2018 ◽  
Vol 64 (12) ◽  
pp. 1704-1712 ◽  
Author(s):  
Felix Neumann ◽  
Iván Hernández-Neuta ◽  
Malin Grabbe ◽  
Narayanan Madaboosi ◽  
Jan Albert ◽  
...  
Keyword(s):  
Seasonal Influenza ◽  
Rolling Circle Amplification ◽  
High Specificity ◽  
Clinical Samples ◽  
Padlock Probe ◽  
Rolling Circle ◽  
Diagnostic Assays ◽  
Padlock Probes ◽  
Probe Assay ◽  
Digital Quantification

Abstract BACKGROUND Influenza remains a constant threat worldwide, and WHO estimates that it affects 5% to 15% of the global population each season, with an associated 3 to 5 million severe cases and up to 500000 deaths. To limit the morbidity and the economic burden of influenza, improved diagnostic assays are needed. METHODS We developed a multiplexed assay for the detection and subtyping of seasonal influenza based on padlock probes and rolling circle amplification. The assay simultaneously targets all 8 genome segments of the 4 circulating influenza variants—A(H1N1), A(H3N2), B/Yamagata, and B/Victoria—and was combined with a prototype cartridge for inexpensive digital quantification. Characterized virus isolates and patient nasopharyngeal swabs were used for assay design and analytical validation. The diagnostic performance was assessed by blinded testing of 50 clinical samples analyzed in parallel with a commercial influenza assay, Simplexa™ Flu A/B & RSV Direct. RESULTS The assay had a detection limit of 18 viral RNA copies and achieved 100% analytical and clinical specificity for differential detection and subtyping of seasonal circulating influenza variants. The diagnostic sensitivity on the 50 clinical samples was 77.5% for detecting influenza and up to 73% for subtyping seasonal variants. CONCLUSIONS We have presented a proof-of-concept padlock probe assay combined with an inexpensive digital readout for the detection and subtyping of seasonal influenza strains A and B. The demonstrated high specificity and multiplexing capability, together with the digital quantification, established the assay as a promising diagnostic tool for seasonal influenza.

AID TB resistance line probe assay for rapid detection of resistant Mycobacterium tuberculosis in clinical samples

2015 ◽  
Vol 70 (4) ◽  
pp. 400-408 ◽  
Author(s):  
B. Molina-Moya ◽  
A. Lacoma ◽  
C. Prat ◽  
J. Diaz ◽  
A. Dudnyk ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Rapid Detection ◽  
Clinical Samples ◽  
Line Probe Assay ◽  
Probe Assay

scholarly journals Comparison of Two Assays for Molecular Determination of Rifampin Resistance in Clinical Samples from Patients with Buruli Ulcer Disease

2014 ◽  
Vol 52 (4) ◽  
pp. 1246-1249 ◽  
Author(s):  
M. Jansson ◽  
M. Beissner ◽  
R. O. Phillips ◽  
K. Badziklou ◽  
E. Piten ◽  
...  
Keyword(s):  
Buruli Ulcer ◽  
Clinical Samples ◽  
Ulcer Disease ◽  
Rifampin Resistance ◽  
Molecular Determination

scholarly journals Characterization of rpoB Mutations in Rifampin-Resistant Clinical Mycobacterium tuberculosisIsolates from Greece

1998 ◽  
Vol 36 (1) ◽  
pp. 20-23 ◽  
Author(s):  
Peggy Matsiota-Bernard ◽  
Georgia Vrioni ◽  
Evangelos Marinis
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Gene Mutations ◽  
Epidemiological Studies ◽  
Dna Sequence Analysis ◽  
Line Probe Assay ◽  
Negative Results ◽  
Pcr Products ◽  
Rifampin Resistance ◽  
Probe Assay

There is a geographic distribution of Mycobacterium tuberculosis strains with various rpoB gene mutations that account for rifampin resistance. We studied 17 rifampin-resistant clinical isolates from patients in Greece to identify rpoBmutations. The aim of our study was the evaluation of a commercially available line probe assay kit (INNO-LiPA Rif. TB) to detectrpoB mutations and rifampin resistance. The results obtained with the commercially available assay were compared to those obtained by automated DNA sequence analysis of amplified PCR products. Randomly amplified polymorphic DNA (RAPD) analyses of the isolates were also performed. The overall concordance of the line probe assay with phenotypic rifampin susceptibility test was 94%. Three distinctrpoB mutations in codons Ser531, His526, and Asp516 were correctly identified with the kit, but mutations in external regions and insertions were detected only by automated DNA sequence analysis. The changes in codons Ser531 and His526 accounted for the majority of rifampin resistance, as previously described for isolates from other geographic areas. The results obtained by RAPD analyses of the isolates suggested that clonally related M. tuberculosis strains can have subclones bearing distinct mutant rpoB alleles. We conclude that this line probe assay kit, which is fast and with which tests are easy to perform, can be used for the rapid detection of rifampin resistance in M. tuberculosis before the availability of results by conventional methods and for epidemiological studies but that negative results obtained by this method do not rule out rifampin resistance.

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