HOW TO ORDER?

How to order?

To order easily and quickly, you can:

Can I be delivered on Saturday?

Products can be delivered on Saturdays in France. Simply add a comment when you order. For other countries, please contact us:

Phone: + 33 (0)9 67 39 35 20
contact@gl-biocontrol.com

How to pay?

Payement has to be done via bank transfer on GL BIOCONTROL account. Other payement methods can be considered but require prior agreement of GL BIOCONTROL.

What if nobody is available to receive a delivery?

The carrier will leave a delivery notice with its contact information.

What is the delivery time of the products?

We do our best to deliver the products within 48 hours in France.

What to do in case of delivery errors?

Delivery errors can be on the product reference or on the quantity. Please, in either case, contact us as soon as possible. Indeed, if products have to be returned, you have a delay of 8 business days (except products to be kept cold).

ATP AND ATP-METRY

What is ATP?

Adenosine triphosphate (ATP) is the major intermediary energy required in most cellular metabolism reactions. Every living cell produces and consumes ATP. This coenzyme, specific to living environments, proves the existence of living organisms.

Which microorganisms are lysed by the DENDRIDIAG® reagent?

DENDRIDIAG® kits preferentially lyses bacteria, cyanobacteria and amoeba. For total lysis of all microorganisms (fungi, yeast, algae…) consult GL BIOCONTROL.

How and how long can I keep the plastic consumables?

Plastic consumables must be stored in a dry area at room temperature. Their expiration date is displayed on their individual packaging.

What do I measure with ATP-metry?

Quantifying ATP equates to quantifying total microorganisms (or total flora) of a sample. ATP-metry is a biomolecular technique, based on bioluminescence. The measurement is done using a luminometer.

At what temperature should I use the reagents?

To ensure a maximal enzyme efficiency, DENDRIDIAG® reagent and STANDARD 1000 must be used at room temperature (18°C – 25°C).

I forgot the DENDRIDIAG® reagent at room temperature. What should I do?

For good stability of the kit, all the reagents must be stored in a freezer (-18°C). If you forgot the reagents at room temperature for a few days, you can perform a control of the reagent efficiency. To do so, refer to the paragraph Control of the reagent efficiency in the handbook’s kit.

What do I measure with the DENDRIDIAG® kits?

With DENDRIDIAG® kits and filtration, only intracellular ATP is measured. It corresponds to the ATP found inside the living cells representative of living bacteria.
Extracellular ATP is also found in sample as a free molecule in the sample. It comes from dead or dying microorganisms. Filtration eliminates free ATP. Without filtration, total ATP is measured: intracellular and extracellular ATP.

How and how long can I store the reagents?

All ATP-metry dropper bottles (DENDRIDIAG® reagents, STANDARD 1000 and EXTRACTANT) must be stored in the dark in a freezer (-18°C). In this way, they can be kept for at least 12 months. After first use, the reagents will be preferentially refrozen. If not, they can be refrigerated (between 3 and 8°C) for a maximum of 8 consecutive weeks. Stored at room temperature, the reagents are stable less than one week.

In which areas of application can use the DENDRIDIAG® kits?

Industrial Water (IW): cooling system, process water circuit, water supply system for industrial purposes…

Sanitary Water (SW): drinking water supply system, water network of spa facilities…

Ultra-Pure Water (UPW): water loop system for medical, pharmaceutical or microelectronics use, water networks under microbiological control…

Surface (BF): swimming pools, food processing, cooling tower, water supply system…

Air (AIR): aeraulic network, hospitals, offices, high risks industries like composting, methanation, farming…

DENDRIDIAG® IW, SW AND UPW KITS

What volume of sample should I filter?

By default, we advise you to filter:
10 ml for the IW kit
50 ml for the SW kit
100 ml for the UPW kit
It is necessary to filter a representative volume of sample in order to get a better reliability of the results.

Anyway, always write down the volume filtered for each sample.

I cannot filter all the sample.

If the sample is highly contaminated, it is possible to clog the filter.

  • If you managed to filter at least 10% of the sample: write down the volume filtered, unscrew the filter and empty the syringe. Put the piston back in the syringe placing the black Teflon part at 3 ml if you use a 10 ml syringe or at 10 ml if you use a 50 ml syringe. Screw the filter back on the syringe and follow the classical protocol.
  • If you did not manage to filter the sample: pour only 1 ml of the sample in the syringe and complete to 10 ml with sterile water if you use a 10 ml syringe or, pour 5 ml of the sample in the syringe and complete to 50 ml with sterile water if you use a 50 ml syringe.

Consult GL BIOCONTROL for further information.

What reference limits should I use for my water analysis?

Based on our experiment, we established the following warning and alarm thresholds:

  • Drinking water production system (in LOG eq.bact./ml):

  • Drinking water supply network (in LOG eq.bact./ml):

  • Domestic hot water – Makeup water (in LOG eq.bact./ml):

  • Process water – Cooling tower water (in LOG eq.bact./ml):


These thresholds should be refined based on the first results obtained on your network.

I cannot get the foam out.

After DENDRIDIAG® suck up, it is necessary to push all the sample out of the syringe, until a white foam comes out. To get the foam out on a 50 ml syringe, apply a strong and constant pressure of few seconds on the piston with the palm of the hand.

If you continue to have difficulties, we can provide you with 10 ml syringes.

Which unit should I use to express my results?

Picogram ATP per milliliter (pgATP/ml) is the real unit.

For a better understanding of the results, it is possible to use the unit equivalent bacteria per milliliter (eq.bact./ml) using the scientific consensus 1 pgATP ≈ 1 000 bacteria. This result is not rigorously true because the ATP concentration varies from microorganism to microorganism and also differs following the metabolic state of the bacteria.

Why is the R2 value inferior to the R1 value?

Signal loss was estimated to 6% to 8% per minute. If measurement takes a long time and there are several minutes between ATP extraction and reading of R2, standardization will happen during the signal degrowth phase. It is often observed on sample highly contaminated.

We advise you to restart the analysis by filtrating a smaller volume of water.

Too much foam in the test tube.

To recover all the microorganisms in the test tube, it is necessary to push all the sample out of the syringe, until a white foam comes out. However, the pressure on the piston must be stopped as soon as the foam comes out in order to avoid formation of a “plug” between the top and the bottom of the tube. During manipulation, lean the test tube so the reagent runs along the tube wall.

If you have an excess of foam or a bubble on the upper part of the test tube, tap the bottom of the tube on a flat surface.

Why express my results in LOG?

Results are often expressed in LOG to simplify the interpretation. Indeed, we consider that two results are significantly different if there is a difference of 1 LOG between the two values.

The following table shows you the conversion of eq.bact./ml in LOG:

The Excel table and the WebApp  DENDRIDIAG®SOFTWARE automatically give you this value.

DENDRIDIAG® BF KIT

What surface dimension should I sample?

Our kit is supplied with a sampling template which defines a surface of 20 cm². With this sampling template, you get a good reproducibility and a significant sampling of the surface studied which leads to reliable results. Nevertheless, if you want to increase the measurement sensitivity, you can perform two sampling of the same surface by moving the sampling template.

Always write down the surface sampled.

What reference limits should I use for my surface analysis?

Based on our experiment, we established the following warning and alarm thresholds:

  • Pool opening (in LOG/cm²):

  • Operating swimming-pools (in LOG/cm²):

  • Surfaces of water pipes (in pgATP/cm²):

These thresholds should be refined based on the first results obtained on your network.

Which unit should I use to express my results?

Picogram ATP per square centimeter (pgATP/cm²) is the real unit.

For a better understanding of the results, it is possible to use the unit equivalent bacteria per square centimeter (eq.bact./cm²) using the scientific consensus 1 pgATP ≈ 1 000 bacteria. This result is not rigorously true because the ATP concentration varies from microorganism to microorganism and also differs following the metabolic state of the bacteria.

Why express my results in LOG?

Results are often expressed in LOG to simplify the interpretation. Indeed, we consider that two results are significantly different if there is a difference of 1 LOG between the two values.

The following table shows you the conversion of eq.bact./cm² in LOG:

The Excel table automatically gives you this value.

qPCR ANALYSIS of LEGIONELLA

What is qPCR?

qPCR means Quantitative Polymerase Chain Reaction. It is a laboratory technique of molecular biology that monitors the amplification of a targeted DNA molecule. It is used to specifically and rapidly quantify microorganisms.  qPCR is already the subject of several ISO standards in the environment and health fields. It is based on the amplification and detection of specific genomic sequences.

Which kit should I use for Legionella analysis by qPCR?

In order to quantify Legionella by qPCR, it is necessary to use a DNA extraction and purification kit, such as the DNA PURE-FLASH kit, and a PCR kit, such as the DNA AMPLI-FLASH Legionella spp and/or Legionella pneumophila kit.

A thermocycler is also required. We offer for sale the thermocycler MIC.

We can also perform the analyses in our laboratory in Montpellier, France. The results are available in under 48 hours after sample receipt. If needed, we can help you to interpret the results. 

What are « Legionella spp » and « Legionella pneumophila » ?

Legionella spp means Legionella species, it means all the species of the genus Legionella. To this day, 58 species have been identified.

Legionella pneumophila is one of the 58 species. Legionella pneumophila is composed of 15 different serovars. It is the species the most sought after for regulatory analyses (e.g. cooling tower circuits, domestic hot water…). Regulation authorizes research of  Legionella pneumophila by qPCR in self-testing.

GL BIOCONTROL can perform these analyses for you.

Does qPCR measure dead bacteria?

No. qPCR technique quantifies the living Legionella present in a water sample. The protocol associated to our DNA extraction and purification kit, DNA PURE-FLASH, includes a filtration step which eliminates free DNA that comes from dead bacteria. The PCR analysis is performed on viable microorganisms that were concentrated on the membrane. In this way, qPCR quantifies only intracellular ATP.

What do the kits DNA AMPLI-FLASH Legionella spp and pneumophila measure?

After DNA extraction and purification with the DNA PURE-FLASH kit, the DNA extract is analyzed by qPCR with the DNA AMPLI-FLASH kit, to measure the genome unit (GU) quantity.

The qPCR analysis directly counts the number of Legionella spp and/or Legionella pneumophila present in the sample. 

DNA PURE-FLASH KIT

How and how long can I store the reagents?

The reagents  DNA PURE-FLASH  can be stored in the fridge (around 4°C) for one year.

The « gel loading » pipette tip got clogged when removing the supernantant.

Remove the tapered tip and discard. Restart the step with a new tip ; they are supplied in excess.

Besides, if there is a lot of WCX resin on your tapered tip, remove the excess of resin by wiping the tip on the internal wall of the tube. 

Resin from the lysis buffer S1 was sampled.

In order to avoid inhibition, it is important to avoid pipetting resin at this step. If a fraction of the resin from the lysis solution (S1) is removed, re-deposit the supernatant in the tube and centrifuge again. 

WCX resin was sampled after the elution step.

In order to avoid inhibition, it is important to avoid pipetting resin at this step. If a fraction of the WCX resin is removed, re-deposit the supernatant in the tube and centrifuge again.

I added 3 drops Binding buffer (S2) instead of 2.

If you had one or two additional drops by mistake, the performance of the protocol will not be impacted. You can continue the purification protocol as you usually would. 

DNA AMPLI-FLASH KITS

How and how long can I store the reagents?

The reagents  DNA AMPLI-FLASH  must be stored in the freezer (around -18°C). In these conditions, they are stable for 6 months.

The complete methodology negative control is positive.

The results cannot be validated. Make sure that the elution buffer is not contaminated. To do so, test it directly in qPCR.

The target (FAM channel) and the internal control (HEX channel) are both negative.

Please, refer to the table page 6 of the document: DNA AMPLI-FLASH – Detailed protocol for quantification of Legionella by qPCR.

I forgot the diluted standard range at room temperature.

Discard all the tubes. From the QS5 solution stored in the freezer, prepare a new calibration range.

The positive control is negative, no wells are positive.

A problem occurs while preparing the mix. Restart the qPCR analysis from the purified samples.

The slope of the calibration range is less than -4.1.

Make sure that the standard range was stored in the freezer. If yes, verify the slope of the calibration range from QS4 to QS2.

  • If the value of the slope is good (> -4.1), it means that the QS1 has degraded.  For the next PCR run, prepare a new dilution of the QS2. Discard the old QS1.
  • If the value of the slope is not good (< -4.1), prepare a new calibration range from the standard solution QS5.

The PCR negative control is positive.

The results can not be validated. Perform a new qPCR analysis from the purified DNA extract.

The internal control (channel HEX) shows no Ct but the target (channel FAM) is positive.

Please, refer to the table page 6 of the document: DNA AMPLI-FLASH – Detailed protocol for quantification of Legionella by qPCR.

ZETTA+ MEMBRANES

What are the ZETTA+ membranes for?

ZETTA+ membranes offer an optimal concentration of bacteriophage and enteroviruses present in sample water. These filters are used for water analyses according to norms ISO 15216-2 and ISO 10705-3.

What analysis method can I use after concentration of viruses on the ZETTA+ membranes?

After viruses concentration on ZETTA+ membranes, analysis can be done by either cultural method or molecular biology methods, such as qPCR or RT-qPCR.

In which application fields can I use the ZETTA+ membranes?

ZETTA+ membranes can be used in various application fields: research of enteroviruses or bacteriophages in bottled water, or drinking water, or irrigation water for cultivation…

What equipment do I need for using ZETTA+ membranes?

It is necessary to have a filtering manifold for 47 mm membranes, and a vacuum pump with the following minimum specifications: 1.2 m3/h – 1.5 mbar. The volume of filtered sample must be adjusted according to the type of water analyzed.

Which norms the ZETTA+ membranes are compatible with?

Thanks to their high retention capacities, the ZETTA+ membranes are compatible with the norms:

  • ISO 15216-2 for research of bacteriophages in water,
  • ISO 10705-3 for research of hepatitis A virus and norovirus in food industry.

LUMINOMETER

My tube holder for PD30 luminometer is dirty. How to clean it?

An excess of foam or a bubble in the upper part of the tube can dirty the tube holder. In this case, you can use a paper towel impregnated with alcohol 70% of water to clean the tube holder.

The luminometer C110 displays « OVERSCALE ».

Luminometer C110 is very sensitive. The message “OVERSCALE” is displayed when the sample is strongly contaminated and the luminometer cannot measure the RLU. If you analyze liquids, restart the measurement by filtrating 1/10th of the volume.

I forgot to write down the value of R1 and R2.

It is possible to retrieve the results in RLU in the luminometer. To do so, turn on the luminometer and after the 10 seconds of calibration, press the up arrow to get the last values obtained.

Luminometer PD30 rings and does not complete calibration.

PD30 calibration is done when the luminometer is empty and the cap correctly closed. If the luminometer rings, make sure the tube holder is out and the cap is well closed.

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