HybriDetect

Universal rapid test for detection of a biotin-and FITC-labeled analytes.
For research use only

Item numberMGHD 1
Package size100 Tests
Sample volume100 μl
  





Background Information about GenLine HybriDetect

Background Information about GenLine HybriDetect

Technology

The Milenia GenLine HybriDetect test strip is a lateral flow test capable of detecting products generated by molecular biology methods. These products are usually created by applying amplification methods, such as PCR, LAMP or RPA. In order to detect the amplicons with the HybriDetect test strip, labeled primers must be used in the amplification procedure. These primers are incorporated in the amplicons in each amplification cycle and Milenia GenLine HybriDetect detects the amplicons via the labels. The principle of PCR product detection is shown in the following figure:

 

The detection of amplification products with the Milenia GenLine HybriDetect is very simple

  • The amplification mix is prepared and distributed to PCR-tubes.
  • Samples are added to the tubes, and in the case of PCR the tubes are transferred to a thermocycler, or if isothermal amplifications like LAMP or RPA are performed, the tubes are transfered to an incubator.
  • After completing the amplification, Milenia GenLine HybriDetect test strips are taken out of their container, labeled individually and the amplification product is pipetted onto the sample application area of the strip.
  • Finally, the strips are placed upright in tubes (e.g. wells of a microtiter plate) that contain the assay buffer included in the kit. Results can be visualized after just 5 – 10 minutes.

For documentation purposes, an image of the strips can be taken with the strips fixed on a sheet of paper. The entire procedure is shown in the following figure:

 

 

Test Development

Usually the development process is started by utilizing agarose gel electrophoresis for the detection of the amplification products. Agarose gels are frequently used in research laboratories to detect nucleic acids via their ability to separate the products according to their molecular weight. The running of agarose gels requires a certain amount of technical knowledge, is time consuming and uses valuable lab resources. In cases when rapid and sensitive tests for individual samples are required by the user, an alternative method is needed.

Milenia GenLine HybriDetect is such an alternative to agarose gels for the detection of amplification products. The benefits of the test strip are as follows:

  • Simple handling
  • Rapid results – 5-10 minutes incubation time
  • High-sensitivity – 100-1000 x more sensitive than an agarose gel
  • Ideal to run one or several samples
  • Strips and Assay Buffer supplied ready to use with the kit – no preparation of reagents necessary
  • Only pipets and a pen are required – no instrumentation needed
  • No ethidium bromide or other harmful compounds in the kit

A detailed comparison of the features of an agarose gel and the HybriDetect test strip is available here.

Possible uses of GenLine HybriDetect

Possible uses of GenLine HybriDetect

Milenia GenLine HybriDetect kits can be used for all types of molecular biological detection, for example, detecting mutations in genes, detecting bacteria and viruses, and also determining specific species. A selection of publications are available here.

At Milenia Biotec, we also use the Milenia GenLine HybriDetect test strip to develop our own PCR evidence and would like to show you some examples:

A Simple PCR-Test for the Detection of Beer Spoilage Organisms

Beer proves a difficult medium for microorganisms to thrive in and multiply due to the presence of alcohol and the low pH. Furthermore, the influence of carbon dioxide and bitter acids from the hops present additional growth barriers. Nevertheless, some microorganisms have adapted to this inhospitable beer environment and are even capable of thriving in such a setting. Unfortunately, this microorganism growth has the potential to spoil the beer, by altering the taste or making it cloudy. For this reason, breweries quality-check their beer, raw materials and the hygiene status of the filling machine by testing for the presence of beer-spoiling organisms at regular intervals throughout the entire production lifecycle. Traditional methods like cell culture take a long time to perform and require expert know-how . The same is true for microscopic procedures. New molecular biology methods, such as real time PCR, are very good in terms of quality. However, they require a high-level of technical ability regarding: test handling, interpretation of the results, and have high instrumentation costs. Therefore, there is a demand for quick, simple-to-handle and reliable tests for the detection of beer-spoilage bacteria, which can be applied in any brewery.

This topic was addressed in a joint research project by Dr. Mathias Hutzler and Jennifer Koob of the Research Center Weihenstephan for Brewing and Food Quality at the TUM , the group of Professor Erdmuthe Meyer zu Bexten at the THM Gießen and Milenia Biotec. For her work on this project, Jennifer Koob was awarded in 2015 by the German Brewing Association the Henrich-Funke-Pschorr-Award

The Milenia GenLine product line for the detection of beer-spoilage organisms can be applied by any brewer within the quality-control methodology of the beer and the monitoring of the hygiene status of the filling machine. The tests are very simple to handle and allow users, after only 2 hours of training and no prior knowledge of molecular biology methods, to perform the tests. A video showing the handling of the tests is available here.

The following tests for the detection of beer spoilage organisms are available:

 

Milenia GenLine Tests Detect Contaminations of Alicyclobacillus in Juice and Help in the Identification of Spoilers Capable of Guajacol Synthesis

Bacteria of the Alicyclobacillus group are gram positive, acidophilic organisms which create spores during their growth. They are well known to spoil juice, smoothies and soft drinks.

Alicyclobacillus cells proliferate at low pH and survive pasteurization. The pasteurization process initiates the formation of spores of the Alicyclobacillus cells and therefore initiates the growth of the cell population. The vegetative cells are capable of spoiling the product. The typical sign of spoilage is the change in taste and the cloudiness of the product. Some Alicyclobacillus species are able to create guajacol, a compound that causes severe changes in taste. The most frequent spoiler of the Alicyclobacillus group is A. acidoterrestris.

The safe identification of Alicyclobacillus in juice is very time consuming if classical cultural methods are applied. According to the guidelines of the IFU (International Fruit Juice Union), the safe identification of Alicyclobacillus contaminatants in juice and other beverages can take up to 15 days. For this reason, rapid and safe identification is highly sought after by many customers. In this respect, a sensitive and precise PCR detection method could dramatically improve the identification of Alicyclobacillus contaminants.

The Milenia GenLine Alicyclobacillus-tests allow the sensitive and specific detection of the Alicyclobacillus group (Alicyclobacillus spec.). If this screening test shows positive results, a second test is available which allows the identification of Alicyclobacillus acidoterrestris. A test for the identification of the capability of the cells to form guajacol is also an integral part of this test. Consequently, this test allows the identification of Alicyclobacillus cells which are cabable of spoiling the product.

The following tests for the identification of Alicyclobacillus are available:

GenLine HybriDetect in the Literature

GenLine HybriDetect in the Literature

RPA

LAMP

PCR

 

 

More about GenLine HybriDetect on milenia-biotec.de

More about GenLine HybriDetect on milenia-biotec.de

Milenia GenLine-Tests for the detection of beer spoilage organisms

Milenia GenLine-Tests for the detection of juice spoilage Alicyclobacillus

Milenia GenLine-Tests under Development

  • Milenia GenLine Mycoplasma in Tissue Culture
  • Milenia GenLine Yeast Screen

Frequently Asked Questions

Frequently Asked Questions

Q:  Is it possible to use FAM instead of FITC as a label of the primers or probes?

A:  Yes, the cross reactivity of the Anti-FITC-Antibody with FAM is very high.

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Q:  When I apply Milenia GenLine HybriDetect in my method, I observe positive results with the strip while I see negative results when I run these samples on an agarose gel. What is the reason for this phenomenon?

A:  Amplificates are separated on an agarose gel according to their molecular weight. Free primers or primer dimers are running in the front of the gel. Milenia HybriDetect detects complexes carrying incorporated labels of either biotin or Digoxigenin and FITC. If primers applied in the amplification procedure bind to each other and therefore form primer dimers, then positive test results occur on the strip! To prevent this from happening, primers and probes must be redesigned in order to prevent the dimerization.