scholarly journals Biosensors: CRISPR/Cas13a‐Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification‐Free miRNA Diagnostics (Adv. Mater. 51/2019)

2019 ◽  
Vol 31 (51) ◽  
pp. 1970365 ◽  
Author(s):  
Richard Bruch ◽  
Julia Baaske ◽  
Claire Chatelle ◽  
Mailin Meirich ◽  
Sibylle Madlener ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Amplification ◽  
Microfluidic Biosensor

scholarly journals CRISPR/Cas13a powered electrochemical microfluidic biosensor for nucleic acid amplification-free miRNA diagnostics

2019 ◽  
Author(s):  
Richard Bruch ◽  
Julia Baaske ◽  
Claire Chatelle ◽  
Mailin Meirich ◽  
Sibylle Madlener ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Limit Of Detection ◽  
Early Stage ◽  
Low Cost ◽  
Clinical Diagnostics ◽  
Nucleic Acid Amplification ◽  
Process Time ◽  
Serum Samples ◽  
Microfluidic Biosensor ◽  
Pcr Method

Non-coding small RNAs, such as microRNAs, are becoming the biomarkers of choice for multiple diseases in clinical diagnostics. A dysregulation of these microRNAs can be associated to many different diseases, such as cancer, dementia or cardiovascular conditions. The key for an effective treatment is an accurate initial diagnosis at an early stage, improving the patient’s survival chances. Here, we introduce a CRISPR/Cas13a powered microfluidic, integrated electrochemical biosensor for the on-site detection of microRNAs. Through this unique combination, the quantification of the potential tumor markers microRNA miR-19b and miR-20a has been realized without any nucleic acid amplification. With a readout time of 9 minutes and an overall process time of less than 4 hours, a limit of detection of 10 pM was achieved, using a measuring volume of less than 0.6 µl. Furthermore, we demonstrate the feasibility of our versatile sensor platform to detect miR-19b in serum samples of children, suffering from brain cancer. The validation of our results with a standard qRT-PCR method shows the ability of our system to be a low-cost and target amplification-free tool for nucleic acid based diagnostics.

scholarly journals CRISPR/Cas13a‐Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification‐Free miRNA Diagnostics

2019 ◽  
Vol 31 (51) ◽  
pp. 1905311 ◽  
Author(s):  
Richard Bruch ◽  
Julia Baaske ◽  
Claire Chatelle ◽  
Mailin Meirich ◽  
Sibylle Madlener ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Amplification ◽  
Microfluidic Biosensor

scholarly journals Ultrastaging Using Ex Vivo Sentinel Lymph Node Mapping and One-Step Nucleic Acid Amplification (OSNA) in Gastric Cancer: Experiences of a European Center

Cancers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 2683
Author(s):  
Bruno Märkl ◽  
Bianca Grosser ◽  
Kerstin Bauer ◽  
Dmytro Vlasenko ◽  
Gerhard Schenkirsch ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Detection Rate ◽  
Ex Vivo ◽  
Sentinel Lymph Node Mapping ◽  
Nucleic Acid Amplification ◽  
Lymph Node Mapping ◽  
Single Positive ◽  
Conventional Histology ◽  
One Step ◽  
Sensitivity Specificity

Background: In this study, the effectiveness of One-step nucleic acid amplification (OSNA) in combination with ex vivo SLN mapping is compared with conventional histology including immunohistochemistry. Methods: LNs were retrieved from gastrectomy specimens in an unfixed state. After ex vivo SLN mapping using methylene-blue, LNs were sliced to provide samples for histology and OSNA. Results: In total, 334 LNs were retrieved in the fresh state from 41 patients. SLN detection was intended in 40 cases but was successful in only 29, with a correct LN status prediction in 23 cases (79%). Excluding one case out of 41 with a failure likely caused by a processing error, OSNA showed a high effectiveness with sensitivity, specificity, and accuracy rates of 85.4%, 93.5%, and 92.4%, respectively. The LN status could be predicted in all but one case, in which the single positive LN was not eligible for OSNA testing. Moreover, OSNA evaluation led to upstaging from N0 to N+ in three cases (14%). Conclusion: The ex vivo SLN protocol used resulted in a relatively poor detection rate. However, the OSNA method was not hampered by this detection rate and proved its potential to increase the sensitivity of metastases detection.

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