Devel­opment of a nucleic acid based COVID-​​19 Rapid Test

Amplicon detection via the lateral flow test Milenia “HybriDetect”

Recently Milenia Biotec GmbH received several customer inquiries related to COVID-​​19 detection, due to the fact, that HybriDetect may be used for the detection of RPA, LAMP or PCR amplicons. Following up these inquiries, we did a liter­ature search on the SHERLOCK (Specific High Sensi­tivity Enzymatic Reporter UnLOCKing) technique, which is a powerful tool to detect genetic infor­mation coming from viruses using CRISPR.


Feluda Test – A new rapid test method for the identi­fi­cation of SARS-​​CoV-​​2

CRISPR-​​test gets approval in India

Just recently, the TATA Group received approval for the commercial launch of “Feluda” (acronym for FNCas9 Editor Limited Uniform Detection Assay) by the Drugs Controller Gerneral of India.

What is Feluda?

The Feluda test is the first CRISPR/​Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats) rapid test available for COVID-​​19 detection. With this new qPCR independent point-​​of-​​care diagnostic approach, the genomic sequence of SARS-​​CoV-​​2 can be detected by Milenias lateral flow assay HybriDetect.


How to overcome conta­m­i­nation in DNA ampli­fi­cation based Lateral Flow Assays

Conta­m­i­nation can happen, no matter what type of ampli­fi­cation method used: LAMP, RPA or PCR

Milenias HybriDetect is a ready-​​to-​​use, universal dipstick based on the lateral flow technology for the detection of various molecules. It is mainly used for the detection of amplicons, generated by LAMP, RPA or PCR by our customers.

As in standard PCR exper­i­ments, which are analyzed by gel electrophoresis or qPCR exper­i­ments, cross-​​contamination is a problem our customers are faced with. Therefore we want to give some infor­mation and advice to help you avoid conta­m­i­na­tions in RPA, LAMP or PCR assays combined with our Lateral Flow tests HybriDetect and HybriDetect 2T.


Milenia HybriDetect Citation List Update July 2020: What are the figures behind the list?

The use of Milenia HybriDetect is described in 129 scien­tific publi­ca­tions – However, there are a lot of inter­esting details behind this number

In 2006 Piepenburg et al. published a paper with the title “DNA detection using recom­bi­nation Proteins” in PLoS, describing the invention of the RPA technology. It was also the very first paper citing the use of Milenia HybriDetect.


A customers question: What is the limit of detection of Milenia HybriDetect?

Milenia HybriDetect is a tool for our customers and us to develop lateral flow based point of care tests.

Test Principle

The principle works as such that biotin and FITC and – if applicable – digox­i­genin and FITC labeled compounds are intro­duced into the test mixture. These compounds form complexes, which are finally detected by the universal lateral flow strips, Milenia HybriDetect.

Since the market intro­duction in the year 2003 a high number of immunoassays and molecular biology method based tests have been developed by researchers. At the time of writing this article in July 2020 we could find 129 scien­tific papers citing the use of Milenia HybriDetect.


A Tool for Mouse Monoclonal Antibody Isotype, Subtype and light Chain Detection

The Milenia Quick Line Mouse Isotyping Kit is designed for fast and easy identi­fi­cation of mouse antibody isotypes, subtypes and light chains and offers numerous appli­ca­tions

Hybridoma Technology

Specific mouse monoclonal antibodies can be produced by the Hybridoma Technology. This method results in immortal hybrid cells formed by B-​​lymphocytes and myeloma cells. After the main steps (immunization of the host animal, B-​​cell and myeloma cell fusion) the exper­i­menter needs to know which subclass is produced by the cells in order to select an antibody clone with the desired features.


Improving the sensi­tivity of the HybriDetect 2T

How to use block oligos for better results

Detecting various PCR amplifi­cates with HybriDetect 2T

The ampli­fi­cation of nucleic acids has become an important tool for various areas such as medical diagnostics, food safety and research over the past decades. A quick and easy way to get a visible result of the ampli­fi­cation is to evaluate the ampli­fi­cation product using the HybriDetect 2T strip, a lateral flow evalu­ation strip.


Lateral Flow based post-​​transplantation Monitoring

CRISPR-​​based diagnosis of infection and rejection with the HybriDetect

HybriDetect helps diagnose viral infec­tions and post-​​transplantation Monitoring with SHERLOCK

Kaminski et al. developed a fast and easy test to detect causes of graft loss in blood and urine samples from post-​​transplant patients after kidney trans­plan­tation.

The need for a new method to detect organ rejec­tions is highly given, because current invasive methods can take several days and are connected with high costs. The loss of time can lead to an increased risk for patients after trans­plan­tation.


Lateral Flow Readout for CRISPR/​Cas-​​based detection strategies

Test– and Control-​​line of the HybriDetect

SHERLOCK and DETECTR are combi­na­tions of isothermal ampli­fi­cation and sensitive CRISPR/​Cas-​​based amplicon recog­nition. The compat­i­bility with the rapid, simple and equipment-​​free HybriDetect lateral flow platform is an essential feature of these methods. Inter­pre­tation of the lateral flow strips is very intuitive and easy.


Diagnostic CRISPR Tools and Techniques

Lateral Flow Assays using Genome Editing Tools

What is CRISPR/​Cas?

The CRISPR /​ Cas system acts as an immune system equiv­alent that is anchored in the genome of bacteria and archaea. The under­lying mechanism protects the organism from phage infec­tions by sequence specific destruction of “unknown” nucleic acids. The CRISPR/​Cas-​​system has the ability to learn, remember and adapt. (1, 2)