PaRTI-Seq® is Micronbrane's complete solution for rapid Pathogen Identification. Combining Devin® filter, optimized sample preparation method for next generation sequencing(NGS) and proprietary analytical methods, it provides precise test results within 24 hours It's much faster, more accurate and cost efficient!
Pa thogen R eal T ime I dentification by - Seq uencing ®
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Sample proccesing Devin microbiome enrichment kit
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Library Prep & Sequencing PaRTI-Seq® miLibrary Preparation kit
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Data Analysing & Reporting PaRTI-Cular™ Analysis Software
Accelerated
Affordable
1 % Accurate

How PaRTI-Seq® works

Sample Prep
DNA Extraction
Library prep
Sequencing
Analysis
Report

Learn how PaRTI-Seq® helps to save more than 50% of sequencing cost

Imagine that each circle represents relative cost of mNGS pathogen detection test for ONE patient. Depending on Sequencing Platform, total cost of the test will be different. If you sequence with HiSeq it will be the lowest, but will require to run 200 samples at the same time, not to mention extremely high capital cost of platform itself If reduce number of samples in run to 18, the cost per sample will be the same as running the single sample on smaller platforms like MiSeq

Smaller sequencing solutions like MiSeq make it possible to test just sample of 1 patient but sequencing cost in that case is still extremely high and make it least favorable option for most patients today.

Cost of test using sequencing depends on many factors like capital cost of sequencer, consumables and reagents, quantity of samples in a sequencing run, and etc., so total sequencing cost of pathogen detection can vary ten and even more times!

On the other hand for standard test output requires as many as 20M reads.

PaRTI-Seq® utilizes Devin® filter which allows to deplete up to 95% of host DNA in 5 minutes and thus significantly increases the percentage of microbial sequencing reads so with PaRTI-Seq® output requires only 5 millions, sequencing reads per sample for the same sensitivity!

Extraction of DNA using Devin can reduce total cost of pathogen detection by sequencing by more than a half!

Analysis & Report Sample Prep & DNA Extraction Library prep Sequencing This is how many patients you need to test at once Cost of one sample x200 x18 x1 Other mNGS tests $$$$ $$$$ $ x4 PaRTI-Seq® $

Integration in your mNGS workflow

Sample Prep DNA Extraction Library prep Sequencing Analysis Report • Deplete host DNA interference in just 5 minutes; • Save up to 75% of sequencing cost • Whole blood, plasma, swabs, CSF, BALF, Ascites

Depending on your needs, you can utilize the whole PaRTI-Seq® technology workflow or integrate just part of it.

For optimal sample preparation we recommend to use Devin® filter which allows to deplete host DNA interference more than 95% in just 5 minutes so you can save up to 75% of sequencing cost at the downstream sequencing.

Devin® filter is intended for use with a wide range of body fluids, including whole blood, plasma, swabs etc. We recommend to use Devin® filter immediately after blood or other liquid is drawn from the patient as long-term storage can greatly affect testing results.

For highly efficient DNA extraction, we recommend to use Devin® Enrichment Kit. Each Devin® Enrichment Kit already includes 24 Devin® filters for 24 reactions and can be used in manual or automated protocol.

Zwitterionic interface Ultra-Self- assemble Coating Technology (ZISC Technology)
< 6 hours
12 hours at 2-6°C
don't freeze

Devin® blood fractionation syringe filter exploits the antifouling Zwitterionic coating technology to specifically capture the white blood cells (>99%), while allowing other blood components to flow through the membrane. This device provides fast and cost-effective solution to reduce the presence of human DNA while elevates the proportion of microbial pathogens in human blood.

We recommend to use Devin® filter immediately after blood is drawn as long-term storage of sample can affect accuracy of results.

Maximum storage time for specimen is 6 hours at ambient temperature. If sample is stored at 2-6 °C, storage time can be extended to 12 hours. Please, DO NOT filter frozen blood samples as it can cause clogging.

To start using Devin® filter Draw 3-10 mL of whole blood sample into the syringe. Open the filter package and securely attach the filter to the syringe on the one side then assemble the needle on the other. Hold the assembled syringe and filter vertically. Press down on the syringe plunger and gently push the sample through the filter to a 15 mL /50 mL falcon tube. Store the filtered sample in the vacutainer and use as soon as possible

Sample prep with Devin® filter

400 x g for 15 min at Room Temperature
16000 x g for 15 min at Room Temperature
Collect the precipitation

1. Draw 3-10 mL of whole blood sample into the syringe. Open the filter package and securely attach the filter to the syringe on the one side then assemble the needle on the other. Hold the assembled syringe and filter vertically. Press down on the syringe plunger and gently push the sample through the filter to a 15 mL /50 mL falcon (centrifugation) tube.

2. Centrifuge whole blood sample at 400 x g for 15 min at room temperature. Collect the plasma (upper layer clear phase) and transfer to a new 15ml / 50ml centrifugation tube.

3. Centrifuge plasma at 16000 x g for 15 min at room temperature. sample pellet will precipitation at the bottom of the centrifugation tube

Collect the precipitation and mark “Sample pellet”

Before starting microbial DNA extraction

mask 1. Please wear a mask and disposable gloves when handling
mask 2. Use sterile consumables to avoid nuclease contamination.
mask 3. Avoid eyes, skin and clothing contact with reagents. In case of any contact, flush with flowing water. flush with flowing water.
mask 4. When the temperature is below 20°C, place the reagent plate in an oven (peMeated 42-60°C) for 5 to 10 mins.
mask 5. Reagent solution contains guanidine salt, avoid using bleach of any contact, flush with flowing water. flush with flowing water.
mask 6. Avoid vigorous shaking, in order to avoid excessive formation of foam.
mask 7. Do not expose opened reagents or plates to air. The evaporation would lead to pH change, or influence the extraction effectiveness.
mask 8. Reagents are all colorless and transparent. Colored reagents indicate contamination, please replace with a fresh plate before proceeding.'

The Nucleic Acid Extraction Kit is suitable for isolating bacterial DNA from whole blood, plasma or other body liquids. Extracted nucleic acids can be analyzed by downstream application, such as real-time PCR and/or next-generation sequencing.

Workflow is shown for whole blood as an example, other fluids have similar workflow

Before start DNA extraction, please make sure that you wear a mask and disposable gloves when handling as well use sterile consumables to avoid nuclease contamination. Reagent solution contains guanidine salt, avoid using bleach of any contact, flush with flowing water. Avoid vigorous shaking, in order to avoid excessive formation of foam. Do not expose opened reagents or plates to air. The evaporation would lead to pH change, or influence the extraction effectiveness

All reagents are colorless and transparent. Colored reagents indicate contamination, so you will need to replace with a fresh plate before proceeding.

If the temperature is below 20℃, please first place the reagent plate in an oven (perheated 42-60℃) for 5 to 10 mins.

All the Kit’s components can be safely transported without any specific conditions however for best results we suggest you to follow the storage conditions shown to you right upon receival of the package!

MICROBIAL DNA ENRICHMENT

Step 1

1. Remove aluminum foil from the well carefully to avoid splashing and pipette well #4 buffer homogeneously. Transfer well #4 buffer into a new eppendorf and label MB on the cap.

2. Transfer well #2 and well #3 buffer into a new eppendorf and label WB1 Mix on the cap

3. Transfer well #5 and well #6 buffer into a new eppendorf and label WB2 Mix on the cap.

Eppendorf MB Eppendorf WB1 Mix Eppendorf WB2 Mix Pipette homogeneously and transfer Transfer and mix Well number 1 2 3 4 5 6
Step 2

1. Put the eppendorf MB on the magnetic rack for 30 seconds to absorb magnetic beads, and recover supernatant to eppendorf WB2 Mix.

2. Remove the eppendorf MB from the magnetic rack. Add 500 µl WB1 Mix and vortex 2 seconds, then spin down.

30 sec 2 sec Eppendorf MB Eppendorf WB1 Mix Eppendorf WB2 Mix 500 µl
Step 3

Add 20 µl Lysozyme + 20 µl Proteinase K + 200 µl Incubation Buffer in sample pellet and pellet are resuspended well. React at 60°C for 10 minutes and vortex for 10 seconds per 4 minutes, then spin down after the reaction.

Eppendorf sample 10 sec 200 µl Incubation Buffer + 20 µl Lysozyme + 20 µl Proteinase K 60°C
Step 4

Add 300 µl 95%~100% EtOH. Vortex for 10 seconds and spin down.

Eppendorf sample 10 sec 300 µl 95%~100% EtOH
Step 5

Put the eppendorf MB on the magnetic rack for 30 seconds to absorb magnetic beads, and recover supernatant to eppendorf WB1 Mix.

Remove the eppendorf MB from the magnetic rack and add the whole buffer of the eppendorf Sample.

30 sec Eppendorf MB Eppendorf WB1 Mix Eppendorf sample ~650 µl
Step 6

Vortex eppendorf MB for 5 minutes and spin down. Put the eppendorf MB on the magnetic rack for 90 seconds to absorb magnetic beads and discard supernatant.

Eppendorf MB 5 min 90 sec
Step 7

Add 750 µl WB1 Mix. Vortex for 30 seconds and spin down. Put the eppendorf MB on the magnetic rack for 60 seconds to absorb magnetic beads and discard supernatant.

Eppendorf MB Eppendorf WB1 Mix 60 sec 30 sec 750 µl
Step 8

Add 750 µl WB2 Mix. Vortex for 30 seconds and spin down. Put the eppendorf MB on the magnetic rack for 60 seconds to absorb magnetic beads and discard supernatant. Repeat step 8 one more time.

Eppendorf MB Eppendorf WB2 Mix 60 sec 30 sec 750 µl
Step 9

Open the eppendorf-MB, and heat at 45°C for 10 minutes to dry the magnetic beads.

Eppendorf MB 10 min 45°C
Step 10

Add 50 µl of Elution buffer into Eppendorf MB and let them react at 45°C for 5 minutes, gently pipetting 5 times per minute.

After reaction completes, spin down and put the Eppendorf MB on the magnetic rack for 60 seconds to absorb magnetic beads. Collect the supernatant EB to a new Lobind tube and mark as the bacterial DNA product.

Extracted bacterial DNA product should be stored at -20°C and can be used for further analysis by downstream application, such as real-time PCR and/or next-generation sequencing.

Eppendorf MB Elution buffer DNA Product 5 min 60 sec 50 µl 45°C

Storage conditions

Reagent TypeTransportation ConditionStorageCondition upon receival
Micronbrane PlateRoom TemperatureRoom Temperature
IBRoom TemperatureRoom Temperature
Proteinase KRoom Temperature (no longer than 5 days)4C (functional refrigerator)
LysozymeRoom Temperature (no longer than 5 days)-20C (functional refrigerator)
Devin® filterRoom TemperatureRoom Temperature

Devin filter and DNA enrichment kit reagents can be safely transported and kept at the room temperature (25°C) in a dry place except Proteinase K (can be safely transported at room temperature no longer than 5 days then best stored at 4C) and Lysozyme (can be safely transported at room temperature no longer than 5 days then best stored at -20C)

FAQ