A New Trace Analytic Method for the Determination of Sequence-Specific DNA with Fluorescent Probes

Zeng, Guoping; Ding, Xiang; Cai, Jin-Zhang

doi:10.1080/00032719.2012.691591

Cited by 3 publications

(4 citation statements)

References 15 publications

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“…The F/F0 value of HCV, which was reported that could not be detected by using ordinary nucleic acid fluorescent gel stains in Zeng's articles [12] , [16] , could be detected in our research by all of the three dyes, and their F/F0 values are 17.05 (SYBR Green II), 26.43 (SYBR Gold), and 63.60 (PicoGreen). From the data, we could find that even for HCV detection, all of the F/F0 values of three dyes are higher than H1N1 in previous report [12] . We considered it as a result of S1 nuclease-mediated cleavage of redundant complementary ssDNA and nonspecific hybridization.…”

Section: Resultsmentioning

confidence: 48%

“…The results were also further evidences that the proposed method could specifically detect the target ssDNA even in contaminant-ridden samples. The present detection limits were much lower than that of the reported one (H1N1, 480 pM, and the other two viruses could not be quantitatively detected by common DNA dyes) [12] .Thus, the high sensitivity and specificity of the strategy may fulfill the requirement for the sensitive and quantitative evaluation of most of ssDNAs.…”

Section: Resultsmentioning

confidence: 60%

“…Compared with Zeng's ssDNA detection method using complementary ssDNA without S1 nuclease [12] , the proposed method in this study shows high sensitivity, specificity and universality. It could not only detect H1N1 virus sequence but also CaM and HCV.…”

Section: Resultsmentioning

confidence: 99%

“…Oligonucleotides (T5, T10, T15, T20, T25, T30, A5, A10, A15, A20, A25, A30, C10, G10) were used for this analysis. Samples were hybridized in 100 µL HB (containing 20 µL 0.5 µM target ssDNA HB solution and its complementary ssDNA HB solution twice as much) by incubating at room temperature for 5 min [12] . Then 90 µL S1 nuclease solution (30 U, prepared according to the instruction: 0.1 µL S1 nuclease per 30 µL 1× reaction buffer, purchased from Fermentas, Beijing, China) was added and incubated at room temperature for 30 min.…”

Section: Methodsmentioning

confidence: 99%

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Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects

Zheng

Liu

et al. 2014

PLoS ONE

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The ability to detect sequence-specific single-strand DNA (ssDNA) in complex, contaminant-ridden samples, using a fluorescent method directly without a DNA extraction and PCR step could simplify the detection of pathogens in the field and in the clinic. Here, we have demonstrated a simple label-free sensing strategy to detect ssDNA by employing its complementary ssDNA, S1 nuclease and nucleic acid fluorescent dyes. Upon clearing away redundant complementary ssDNA and possibly mismatched double strand DNA by using S1 nuclease, the fluorescent signal-to-noise ratio could be increased dramatically. It enabled the method to be adaptable to three different types of DNA fluorescent dyes and the ability to detect target ssDNA in complex, multicomponent samples, like tissue homogenate. The method can distinguish a two-base mismatch from avian influenza A (H1N1) virus. Also, it can detect the appearance of 50 pM target ssDNA in 0.5 µg·mL−1 Lambda DNA, and 50 nM target ssDNA in 5 µg·mL−1 Lambda DNA or in tissue homogenate. It is facile and cost-effective, and could be easily extended to detect other ssDNA with many common nucleic acid fluorescent dyes.

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Section: Resultsmentioning

confidence: 48%

Section: Resultsmentioning

confidence: 60%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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