3. Detection of red tide dinoflagellate
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Karenia mikimatoi is a common red tide alga, which can induce the death of fish and shellfish and therefore causes economic losses. The authors developed a new detection method using a LAMP combined with our HybriDetect for visualization. The whole procedure takes only 23 minutes from preparation of the sample to result.

Only a simple water bath or heat block is needed. The LAMP combined with HybriDetect shows is more specific and faster, compared to agarose gel electrophoresis or simple adding of SYBR green. The authors could detect the results 3 minutes after inserting the dipstick into their probe.

Reference:

Huang H. L., Gao W.F., Zhu P., Zhou C.X., Qiao L.L., Dang C.Y., Pang J.H., Yan X.J., Molecular method for rapid detection of the red tide dinoflagellate Karenia mikimotoi in the coastal region of Xiangshan Bay, China. Journal of Microbiological Methods (2020); V 168

4. Cas9-Mediated Lateral Flow Nucleic Acid Assay
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Wang et al. presented a new method called CRISPR/Cas9-mediated lateral flow nucleic acid assay (CASLFA), which is one of the most convenient analytical techniques for analyzing immune response. They identified Listeria monocytogenes, GMOs and African swine fever virus (ASFV) with a detection limit of hundreds of copies of genome samples with high specificity within one hour on our HybriDetect. ASFV infected samples could be detected with an accuracy of 100% compared to RT-PCR.

The authors point out, that no laboratory environment is needed. The point-of-care-use without any technical expertise and ancillary equipment is given with this new technique.

Reference:

Wang X., Xion E., Tian T., Cheng M., Lin W., Wang H. Clustered Regularly Interspaced Short Palindromic Repeats / Cas9- Mediated Lateral Flow Nucleic Acid Assay. ACS Nano (2020), IF 13.903.

5. Detecting Plant Genes
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Abudayyeh et al. presented a modified SHERLOCK detection method to quantify levels of the glyphosate resistance gene in a mixture of soybeans and to detect multiple plant genes in a single reaction. They described a field-ready SHERLOCK platform combined with Milenias HybriDetect for detection of genes in a range of agricultural applications.

Prior the Cas-reaction (20 minutes), a Recombinase Polymerase Amplification (RPA), which takes 10 minutes, is done.

The high sensitivity (single molecule, 2 aM input concentration in 1 µL sample) and specificity for single nucleotide discrimination is the major advantage of this technique. It can be used for field application, and easily combined with HybriDetect when a reporter labeled with biotin and FAM is added to the SHERLOCK reaction.

Taken together the described method is a useful platform for many biotechnological and agricultural applications.

Reference:

Abudayyeh O. O., Gootenberg J. S., Kellner M. J., Zhang F., Nucleic Acid Detection of Plant Genes Using CRISPR-Cas13. The CRISPR Journal (2019), V 2 (3, 165-171)

6. Detecting Pathogens and GMOs
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Limit of rapid, low cost, user-friendly, field deployable DNA test method

Another plant application using CRISPR and HybriDetect is described by Zhang et al.. The group used a Cas12 based DNA detection method for crop disease diagnosis and a GMO test. They also used a RPA combined with Cas12 cleavage and HybriDetect for visualization.

A great advantage of this method is that it runs at 37°C and no extra instruments except filter paper and HybriDetect is needed.

The tested rice blast pathogen and Bt-rice (transgenic rice) were efficiently identified from leaf disc samples.

The described method is a rapid, low-cost, user-friendly DNA test method, which can be easily applied in field for crop disease diagnosis and GMO administration.

Reference:

Zhang Y., Zhang Y., Xie K., Evaluation of CRISPR/Cas12a-based DNA detection for fast pathogen diagnosis and GMO test in rice. Molecular Breeding (2020); V 40,11

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