Semi-automatic instrumentation for nucleic acid extraction and purification to quantify pathogens on surfaces

The Analyst ◽  
2019 ◽  
Vol 144 (22) ◽  
pp. 6586-6594 ◽  
Author(s):  
Won-Nyoung Lee ◽  
Hyun Jin Yoo ◽  
Kim Huyen Nguyen ◽  
Changyoon Baek ◽  
Junhong Min

A semi-automated detection system compatible with PCR that can detect infectious pathogens on wide surfaces in a short time.

The Analyst ◽  
2020 ◽  
Vol 145 (6) ◽  
pp. 2412-2419 ◽  
Author(s):  
Rachel N. Deraney ◽  
Lindsay Schneider ◽  
Anubhav Tripathi

NA extraction and purification utilitzing a microfluidic chip with applied electric field to induce electroosmotic flow opposite the magnetic NA-bound bead mix.


2001 ◽  
Vol 47 (3) ◽  
pp. 378-383 ◽  
Author(s):  
Chieko Matsumoto ◽  
Rieko Shiozawa ◽  
Shigeki Mitsunaga ◽  
Akiko Ichikawa ◽  
Rika Ishiwatari ◽  
...  

PLoS ONE ◽  
2018 ◽  
Vol 13 (7) ◽  
pp. e0199869 ◽  
Author(s):  
Nitu Thakore ◽  
Ryan Norville ◽  
Molly Franke ◽  
Roger Calderon ◽  
Leonid Lecca ◽  
...  

Lab on a Chip ◽  
2019 ◽  
Vol 19 (22) ◽  
pp. 3853-3861 ◽  
Author(s):  
Chenguang Zhang ◽  
Gongchen Sun ◽  
Satyajyoti Senapati ◽  
Hsueh-Chia Chang

We report a new Bifurcated Continuous Field-Flow Fractionation (BCFFF) microfluidic chip for isolation and purification of nucleic acids from blood plasma with high and concentration-independent yield. The platform is ideal for isolation and quantification of small miRNAs.


Micromachines ◽  
2017 ◽  
Vol 8 (7) ◽  
pp. 228 ◽  
Author(s):  
Darren Branch ◽  
Erika Vreeland ◽  
Jamie McClain ◽  
Jaclyn Murton ◽  
Conrad James ◽  
...  

2002 ◽  
Vol 2 (3) ◽  
pp. 95-100 ◽  
Author(s):  
J. Dellundé ◽  
S. Pina ◽  
J. Jofre ◽  
F. Lucena

A new protocol based on a combination of an excystation process followed by a nucleic acid extraction that combines the lysing and nuclease-inactivating properties of the chaotropic agent guanidinium thiocyanate and the nucleic acid-binding properties of silica particles is described for the extraction and purification of nucleic acids from Cryptosporidium. The application of nested and/or semi-nested PCR using different external and internal primers to DNA extracted by this method from seeded and naturally occurring Cryptosporidium oocysts concentrated and purified from environmental samples detects numbers of oocysts ranging from 20 to 50. The method is feasible, detects mostly excystable oocysts and no problems of inhibition of PCR were observed when applied to environmental samples.


Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 188
Author(s):  
Tanja Hoffmann ◽  
Andreas Hahn ◽  
Jaco J. Verweij ◽  
Gérard Leboulle ◽  
Olfert Landt ◽  
...  

This study aimed to assess standard and harsher nucleic acid extraction schemes for diagnostic helminth real-time PCR approaches from stool samples. A standard procedure for nucleic acid extraction from stool and a procedure including bead-beating as well as proteinase K digestion were compared with group-, genus-, and species-specific real-time PCR assays targeting helminths and nonhelminth pathogens in human stool samples. From 25 different in-house and commercial helminth real-time PCR assays applied to 77 stool samples comprising 67 historic samples and 10 external quality assessment scheme samples positively tested for helminths, higher numbers of positive test results were observed after bead-beating-based nucleic acid extraction for 5/25 (20%) real-time PCR assays irrespective of specificity issues. Lower cycle threshold values were observed for one real-time PCR assay after the standard extraction scheme, and for four assays after the bead-beating-based scheme. Agreement between real-time PCR results after both nucleic acid extraction strategies according to Cohen’s kappa ranged from poor to almost perfect for the different assays. Varying agreement was observed in eight nonhelminth real-time PCR assays applied to 67 historic stool samples. The study indicates highly variable effects of harsh nucleic acid extraction approaches depending on the real-time PCR assay used.


Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 615
Author(s):  
Allen Wing-Ho Chu ◽  
Cyril Chik-Yan Yip ◽  
Wan-Mui Chan ◽  
Anthony Chin-Ki Ng ◽  
Dream Lok-Sze Chan ◽  
...  

SARS-CoV-2 RT-PCR with pooled specimens has been implemented during the COVID-19 pandemic as a cost- and manpower-saving strategy for large-scale testing. However, there is a paucity of data on the efficiency of different nucleic acid extraction platforms on pooled specimens. This study compared a novel automated high-throughput liquid-based RNA extraction (LRE) platform (PHASIFYTM) with a widely used magnetic bead-based total nucleic acid extraction (MBTE) platform (NucliSENS® easyMAG®). A total of 60 pools of nasopharyngeal swab and 60 pools of posterior oropharyngeal saliva specimens, each consisting of 1 SARS-CoV-2 positive and 9 SARS-CoV-2 negative specimens, were included for the comparison. Real-time RT-PCR targeting the SARS-CoV-2 RdRp/Hel gene was performed, and GAPDH RT-PCR was used to detect RT-PCR inhibitors. No significant differences were observed in the Ct values and overall RT-PCR positive rates between LRE and MBTE platforms (92.5% (111/120] vs 90% (108/120]), but there was a slightly higher positive rate for LRE (88.3% (53/60]) than MBTE (81.7% (49/60]) among pooled saliva. The automated LRE method is comparable to a standard MBTE method for the detection of SAR-CoV-2 in pooled specimens, providing a suitable alternative automated extraction platform. Furthermore, LRE may be better suited for pooled saliva specimens due to more efficient removal of RT-PCR inhibitors.


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