T7 exo-mediated FRET-breaking combined with DSN–RNAse–TdT for the detection of microRNA with ultrahigh signal-amplification

The Analyst ◽  
2019 ◽  
Vol 144 (10) ◽  
pp. 3216-3220 ◽  
Author(s):  
Van Thang Nguyen ◽  
Binh Huy Le ◽  
Young Jun Seo

A DSN–RNAse–TdT–T7 exo probing system allows the detection of miRNA 21 with very high sensitivity (LOD = 2.57 fM) and selectivity—the result of (i) avoiding the false-positive signal from miRNA reacting with TdT polymerase and (ii) signal amplification occurring through a FRET-breaking mechanism involving T7 exo.

1990 ◽  
Vol 120 (3) ◽  
pp. 698-699 ◽  
Author(s):  
Stanley S. Schrem ◽  
Mark Nachamie ◽  
Edwin Weiss

2018 ◽  
Vol 17 ◽  
Author(s):  
Todd Talarico ◽  
Michael Murphy ◽  
Raymond Nims ◽  
Dan Hastings ◽  
Jeri Ann Boose ◽  
...  

1992 ◽  
Vol 123 (2) ◽  
pp. 412-416 ◽  
Author(s):  
Debra K. Moser ◽  
William G. Stevenson ◽  
Mary A. Woo

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Eiki Yamasaki ◽  
Ryuta Sakamoto ◽  
Takashi Matsumoto ◽  
Fumiki Morimatsu ◽  
Takayuki Kurazono ◽  
...  

Because cholera toxin (CT) is responsible for most of the symptoms induced byVibrio choleraeinfection, detection of CT is critical for diagnosis of the disease. In this study, we constructed an immunochromatographic test strip for detection of CT (CT-IC) with polyclonal antibodies developed against purified recombinant whole CT protein. The detection limit of the CT-IC was 10 ng/mL of purified recombinant CT, and it could detect the CT in culture supernatant of all 15 toxigenicV. choleraeisolates examined, whereas no false-positive signal was detected in all 5 nontoxigenicV. choleraeisolates examined. The specificity of the CT-IC was examined with recombinant heat-labile toxin (LT), which shares high homology with CT, and it was revealed that the minimum detection limit for LT was 100 times higher than that for CT. In addition,ltgene-positive enterotoxigenicEscherichia coli(ETEC) was examined by CT-IC. The false-positive signals were observed in 3 out of 12 ETEC isolates, but these signals were considerably faint. The CT-IC did not develop false-positive signals with all 7V. parahaemolyticusisolates. These results showed the high specificity of CT-IC and the feasible use of it for the detection and surveillance of toxigenicV. cholerae.


1992 ◽  
Vol 123 (6) ◽  
pp. 1701-1703 ◽  
Author(s):  
Eric Taylor ◽  
Mark Effron ◽  
Enrico P. Veltri

Bioanalysis ◽  
2020 ◽  
Vol 12 (15) ◽  
pp. 1071-1085
Author(s):  
Jihua Chen ◽  
Kimberly Kendra ◽  
Albert Torri ◽  
Giane Sumner

Background: Soluble multimeric target proteins can generate a target-mediated false-positive signal in bridging anti-drug antibody (ADA) assays. A high background signal due to target interference was observed in our anti-REGN-Y antibody assay, and two different strategies were evaluated to mitigate this false-positive signal. Results: Multiple anti-target antibodies were tested and found to be ineffective at reducing target interference, so soluble target receptor and co-factor proteins were used in combination to inhibit the target-mediated signal. These competitive blockers synergistically inhibited target interference and increased target tolerance levels, especially when the assay was performed under mild acidic conditions. A separate approach, target immunodepletion using magnetic beads conjugated with an anti-target antibody, was also effective at mitigating the target-mediated signal, also in combination with mild acidic assay pH. Both methods allowed detection of a true ADA signal in monkey and human post-dose serum samples. Conclusion: These methods provide alternative strategies for mitigating target interference when standard anti-target antibodies are ineffective, with the competitive blocker method being recommended, if possible, due to its higher throughput and easier execution.


Author(s):  
Phu C. Tran ◽  
Will DeBrock ◽  
Mary E. Lester ◽  
Brett C. Hartman ◽  
Juan Socas ◽  
...  

Abstract Background Transcutaneous tissue oximetry is widely used as an adjunct for postoperative monitoring after microvascular breast reconstruction. Despite a high sensitivity at detecting vascular issues, alarms from probe malfunctions/errors can generate unnecessary nursing calls, concerns, and evaluations. The purpose of this study is to analyze the false positive rate of transcutaneous tissue oximetry monitoring over the postoperative period and assess changes in its utility over time. Patients and Methods Consecutive patients undergoing microvascular breast reconstruction at our institution with monitoring using transcutaneous tissue oximetry were assessed between 2017 and 2019. Variables of interest were transcutaneous tissue oximetry alarms, flap loss, re-exploration, and salvage rates. Results The study included 175 patients (286 flaps). The flap loss rate was 1.0% (3/286). Twelve patients (6.8%) required re-exploration, with 9 patients found to have actual flap compromise (all within 24 hours). The salvage rate was 67.0%. The 3 takebacks after 24 hours were for bleeding concerns rather than anastomotic problems. Within the initial 24-hour postoperative period, 43 tissue oximetry alarms triggered nursing calls; 7 alarms (16.2%) were confirmed to be for flap issues secondary to vascular compromise. After 24 hours, none of the 44 alarms were associated with flap compromise. The false positive rate within 24 hours was 83.7% (36/43) compared with 100% (44/44) after 24 hours (p = 0.01). Conclusion The transcutaneous tissue oximetry false positive rate significantly rises after 24 hours. The benefit may not outweigh the concerns, labor, and effort that results from alarms after postoperative day 1. We recommend considering discontinuing this monitoring after 24 hours.


Author(s):  
T.Y. Chang ◽  
J.E. Zucker ◽  
K.L. Jones ◽  
N.J. Sauer ◽  
B. Tell ◽  
...  

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