Easy method for discriminating the origins of manila clam Ruditapes philippinarum with a dual-labelled PNA-probe-based melting curve analysis

Kim, Eun Mi; Song, Min Sik; Hur, Deok Hwe; An, Cheul Min; Kang, Jung-Ha; Park, Jung Youn

doi:10.1007/s13206-015-9402-1

Cited by 4 publications

(2 citation statements)

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“…Probe-based fluorescence melting curve analysis is also a powerful tool to detect mutations. 8 PNA probes hybridize with their complementary target DNA sequences, 26 and PNA–DNA hybridization is more sensitive to base mismatches than DNA–DNA hybridization. Because PNA probe-based fluorescence melting curve analysis is based on the temperature at which the probe-target hybridization dissociates, 27 the results are much easier to interpret than those of Sanger sequencing.…”

Section: Discussionmentioning

confidence: 99%

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Peptide Nucleic Acid Probe-Based Analysis as a New Detection Method for Clarithromycin Resistance inHelicobacter pylori

et al. 2018

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Background/AimsHelicobacter pylori eradication rates are decreasing because of increases in clarithromycin resistance. Thus, finding an easy and accurate method of detecting clarithromycin resistance is important.MethodsWe evaluated 70 H. pylori isolates from Korean patients. Dual-labeled peptide nucleic acid (PNA) probes were designed to detect resistance associated with point mutations in 23S ribosomal ribonucleic acid gene domain V (A2142G, A2143G, and T2182C). Data were analyzed by probe-based fluorescence melting curve analysis based on probe-target dissociation temperatures and compared with Sanger sequencing.ResultsAmong 70 H. pylori isolates, 0, 16, and 58 isolates contained A2142G, A2143G, and T2182C mutations, respectively. PNA probe-based analysis exhibited 100.0% positive predictive values for A2142G and A2143G and a 98.3% positive predictive value for T2182C. PNA probe-based analysis results correlated with 98.6% of Sanger sequencing results (κ-value=0.990; standard error, 0.010).ConclusionsH. pylori clarithromycin resistance can be easily and accurately assessed by dual-labeled PNA probe-based melting curve analysis if probes are used based on the appropriate resistance-related mutations. This method is fast, simple, accurate, and adaptable for clinical samples. It may help clinicians choose a precise eradication regimen.

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Section: Discussionmentioning

confidence: 99%

“…In recent years, eradication rates with first-line therapy in South Korea have ranged from 74.6% to 75.8%. 6 – 8 Clarithromycin resistance is due to point mutations in the peptidyl transferase encoding region in domain V of the H. pylori 23S ribosomal ribonucleic acid (rRNA) gene. 9 , 10 …”

Section: Introductionmentioning

confidence: 99%