scholarly journals Locked Nucleic Acid Probe-Based Real-Time PCR Assay for the Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

PLoS ONE ◽  
2015 ◽  
Vol 10 (11) ◽  
pp. e0143444 ◽  
Author(s):  
Yong Zhao ◽  
Guilian Li ◽  
Chongyun Sun ◽  
Chao Li ◽  
Xiaochen Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Locked Nucleic Acid ◽  
Pcr Assay ◽  
Locked Nucleic Acid Probe

scholarly journals Rapid Detection of Acquired and Inducible Clarithromycin Resistance in Mycobacterium abscessus Group by a Simple Real-Time PCR Assay

2015 ◽  
Vol 53 (7) ◽  
pp. 2337-2339 ◽  
Author(s):  
Robert F. Luo ◽  
Cheyenne Curry ◽  
Nathan Taylor ◽  
Indre Budvytiene ◽  
Niaz Banaei
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Mycobacterium Abscessus ◽  
Nucleic Acid Testing ◽  
Pcr Assay ◽  
Content Type ◽  
Clarithromycin Resistance ◽  
Group A

By targeting theerm(41) andrrlgenes in theMycobacterium abscessusgroup, a multiplex real-time PCR assay for clarithromycin resistance showed 95% (38/40) concordance with nucleic acid testing and 95% (37/39) concordance with phenotypic testing. This assay provides a simple and rapid alternative to extended incubation orerm(41) sequencing.

Locked nucleic acid probe-based real-time PCR for the diagnosis of Listeria monocytogenes in ruminants

2016 ◽  
Vol 30 (3) ◽  
pp. 138-145 ◽  
Author(s):  
Mohamed Barkallah ◽  
Yaakoub Gharbi ◽  
Mariam Hmani ◽  
Zouhir Mallek ◽  
Michel Gautier ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Locked Nucleic Acid ◽  
Locked Nucleic Acid Probe

scholarly journals Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chien-Ru Lin ◽  
Hsin-Yao Wang ◽  
Ting-Wei Lin ◽  
Jang-Jih Lu ◽  
Jason Chia-Hsun Hsieh ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Nucleic Acid ◽  
Real Time ◽  
Sensitivity And Specificity ◽  
Real Time Pcr ◽  
Disease Transmission ◽  
High Sensitivity ◽  
Nucleic Acid Amplification ◽  
Amplification Efficiency ◽  
Pcr Assay

AbstractThe Mycobacterium tuberculosis complex (MTBC) remains one of the top 10 leading causes of death globally. The early diagnosis of MTBC can reduce mortality and mitigate disease transmission. However, current nucleic acid amplification diagnostic test methods are generally time-consuming and show suboptimal diagnostic performance, especially in extrapulmonary MTBC samples or acid-fast stain (AFS)-negative cases. Thus, development of an accurate assay for the diagnosis of MTBC is necessary, particularly under the above mentioned conditions. In this study, a single-tube nested real-time PCR assay (N-RTP) was developed and compared with a newly in-house-developed high-sensitivity real-time PCR assay (HS-RTP) using 134 clinical specimens (including 73 pulmonary and 61 extrapulmonary specimens). The amplification efficiency of HS-RTP and N-RTP was 99.8% and 100.7%, respectively. The sensitivity and specificity of HS-RTP and N-RTP for the diagnosis of MTBC in these specimens were 97.5% (77/79) versus 94.9% (75/79) and 80.0% (44/55) versus 89.1% (49/55), respectively. The sensitivity and specificity of HS-RTP and N-RTP for the diagnosis of MTBC in pulmonary specimens were 96.3% (52/54) versus 96.3% (52/54) and 73.7.0% (14/19) versus 89.5% (17/19), respectively; in extrapulmonary specimens, the sensitivity and specificity of HS-RTP and N-RTP were 100% (25/25) versus 92% (23/25) and 83.3% (30/36) versus 88.9% (32/36), respectively. Among the AFS-negative cases, the sensitivity and specificity of HS-RTP and N-RTP were 97.0% (32/33) versus 90.9% (30/33) and 88.0% (44/50) versus 92.0% (46/50), respectively. Overall, the sensitivity of HS-RTP was higher than that of N-RTP, and the performance was not compromised in extrapulmonary specimens and under AFS-negative conditions. In contrast, the specificity of the N-RTP assay was higher than that of the HS-RTP assay in all types of specimens. In conclusion, the HS-RTP assay would be useful for screening patients suspected of exhibiting an MTBC infection due to its higher sensitivity, while the N-RTP assay could be used for confirmation because of its higher specificity. Our results provide a two-step method (screen to confirm) that simultaneously achieves high sensitivity and specificity in the diagnosis of MTBC.

scholarly journals New Real-Time PCR Assay Using Locked Nucleic Acid Probes To Assess Prevalence of ParC Mutations in Fluoroquinolone-Susceptible Streptococcus pneumoniae Isolates from France

2006 ◽  
Vol 50 (4) ◽  
pp. 1594-1598 ◽  
Author(s):  
Jean-Winoc Decousser ◽  
Imen Methlouthi ◽  
Patrick Pina ◽  
Anne Collignon ◽  
Pierre Allouch
Keyword(s):  
Streptococcus Pneumoniae ◽  
Nucleic Acid ◽  
High Risk ◽  
Real Time ◽  
Real Time Pcr ◽  
Locked Nucleic Acid ◽  
Pcr Assay ◽  
Nucleic Acid Probes ◽  
National Surveys ◽  
Risk Patients

ABSTRACT A real-time PCR assay with locked nucleic acid probes was developed to screen mutations at codons 79 and 83 of the Streptococcus pneumoniae parC gene. Only silent mutations were detected among 236 French invasive fluoroquinolone-susceptible strains. This test could be useful for some high-risk patients or in national surveys.

scholarly journals Real-Time Detection of the Mutation Responsible for Progressive Rod-Cone Degeneration in Labrador Retriever Dogs Using Locked Nucleic Acid TaqMan Probes

2009 ◽  
Vol 21 (5) ◽  
pp. 689-692 ◽  
Author(s):  
Fabio Gentilini ◽  
Gian Luca Rovesti ◽  
Maria Elena Turba
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Late Onset ◽  
Labrador Retriever ◽  
Locked Nucleic Acid ◽  
Sensitivity Threshold ◽  
Ophthalmological Examination ◽  
Pcr Assay ◽  
Labrador Retrievers

Progressive rod-cone degeneration ( prcd) is a late onset, autosomal recessive, inherited disease in dogs caused by a G > A substitution in the PRCD locus, which has been reported in more than 18 breeds, including Labrador Retriever dogs. In this study, a real-time polymerase chain reaction (PCR) assay, exploiting the features of locked nucleic acid (LNA) fluorescent-labeled probes, was developed to genotype the sequence variants responsible for the disease. Two Labrador Retrievers were diagnosed with prcd by ophthalmological examination performed by a panelist of the Italian hereditary eye disease control program. The 2 dogs, as well as 8 related and 14 unrelated Labrador Retrievers, were genotyped with both direct sequencing of the disease locus and real-time LNA TaqMan PCR assay. Even though the region surrounding the mutation was predicted to be highly structured, making probe annealing difficult, the real-time PCR assay allowed researchers to correctly genotype the dogs in all cases with a sensitivity threshold of 4 ng/reaction of genomic DNA. A real-time PCR assay will allow a high-throughput analysis of a larger cohort of dogs, thereby enabling researchers to investigate the prevalence of the mutated allele in the affected breeds.

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