Thermo Fisher Scientific MagMAX Dx Prefilled Viral/Pathogen Nucleic Acid Isolation Kit User guide

Brand: Thermo Fisher Scientific

Model: MagMAX Dx Prefilled Viral/Pathogen Nucleic Acid Isolation Kit

Type: User guide

For In Vitro Diagnostic Use.

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic

Acid Isolation Kit

USER GUIDE

Catalog Numbers A52076

Publication Number MAN0025460

Revision B.0

Invitrogen Bioservices Private Ltd. | Citius Block, First Technology Place, No. 3 EPIP | Whiteﬁeld, Bangalore 560066, India

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The customer is responsible for validation of assays and compliance with regulatory requirements that pertain to their procedures and

uses of the instrument.

The information in this guide is subject to change without notice.

DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE

LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR

ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.

Revision history: Pub. No. MAN0025460

Revision Date Description

B.0 01 February 2022 Correcting "Limited product warranty" section to remove phone number.

A.0 10 August 2021 New manual.

Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these

products, you accept the terms and conditions of all applicable Limited Use Label Licenses.

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Contents

■CHAPTER 1 Product information .................................................. 4

Intended use ................................................................... 4

Product information ............................................................. 4

Contents and storage ............................................................ 5

Required materials not supplied ................................................... 5

■CHAPTER 2 Methods ............................................................... 7

Procedural guidelines ............................................................ 7

Guidelines for saliva collection ................................................ 7

Guidelines for oropharyngeal and nasopharyngeal swabs ........................ 8

Before you begin ................................................................ 8

Before each use of the kit .................................................... 8

Warnings and precautions ................................................... 8

Prepare samples for extraction .................................................... 9

Prepare raw saliva samples .................................................. 9

Prepare preserved saliva samples ............................................. 9

Prepare swab samples for extraction .......................................... 9

Perform viral nucleic acid isolation ............................................... 10

Set up the instrument ...................................................... 10

Prepare the Sample Plate, then process samples on the instrument .............. 10

■APPENDIX A Troubleshooting .................................................... 12

■APPENDIX B Safety ............................................................... 14

Chemical safety ................................................................ 14

Biological hazard safety ......................................................... 15

■APPENDIX C Documentation and support ...................................... 16

Customer and technical support ................................................. 16

Limited product warranty ........................................................ 16

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 3

Product information

WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear

appropriate protective eyewear, clothing, and gloves. Safety Data Sheets (SDSs) are available from

thermoﬁsher.com/support.

Intended use

The MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit is a magnetic bead technology-

based nucleic acid puriﬁcation kit intended for isolation and puriﬁcation of viral RNA and DNA from

human saliva, nasopharyngeal and oropharyngeal swabs for the detection of SARS-CoV-2 virus,

adenovirus, and inﬂuenza virus by real-time PCR method.

The kit is intended for use by qualiﬁed and trained clinical laboratory personnel speciﬁcally instructed

and trained in the techniques of automated magnetic bead puriﬁcation and in vitro diagnostic

procedures.

Product information

The Applied Biosystems™ MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit (Cat. No.

A52076) is speciﬁcally designed for scalable, rapid puriﬁcation of high-quality total nucleic acid from

easy-to-lyse virus in saliva or transport media. The kit utilizes MagMAX™ magnetic-bead technology,

ensuring reproducible recovery of high-quality nucleic acid for a range of downstream applications

such as sequencing and qPCR. This protocol guides users through automated isolation of RNA and

DNA from raw and stabilized saliva samples and transport media using the KingFisher™ Flex Magnetic

Particle Processor.

4MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

Contents and storage

Reagents that are provided in each kit support 192 reactions.

IMPORTANT! On receipt, store all plates and reagents in an upright position at room temperature.

Table 1 MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit

Component Quantity Storage

96 deep-well plate ﬁlled with Binding Solution (samples will be added

to this plate) 2

Store upright at

15°C to 25°C.

96 deep-well plate ﬁlled with Magnetic Beads 2

96 deep-well plate ﬁlled with Wash I Solution 2

96 deep-well plate ﬁlled with Wash II Solution 2

96 deep-well plate ﬁlled with Elution Solution 2

96 deep-well tip comb 2

Proteinase K Solution 1 vial

MicroAmp™ Clear Adhesive Film 2

Required materials not supplied

Unless otherwise indicated, all materials are available through thermoﬁsher.com. MLS: Fisher

Scientiﬁc (ﬁsherscientiﬁc.com) or another major laboratory supplier.

Table 2 For bioﬂuid or transport media sample protocols

Item Source

Instrument

KingFisher™ Flex Magnetic Particle Processor 96DW with 96 Deep-well Head 5400630

KingFisher™ Flex 96 Deep-Well Heating Block (do not use a standard heating

block) 24075430

Equipment

Single and multichannel adjustable pipettors (1.00 µL to 1,000.0 µL) MLS

Laboratory mixer, vortex, or equivalent MLS

Cold block or ice MLS

Tubes, plates and other consumables

PBS (1X), pH 7.4 (without calcium and magnesium)[1] 10010023

Chapter 1 Product information

Contents and storage 1

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 5

Table 2 For bioﬂuid or transport media sample protocols (continued)

Item Source

MicroAmp™ Adhesive Film Applicator 4333183

Non-stick, RNase-free microcentrifuge tubes (1.5 mL and 2.0 mL)[2] thermoﬁsher.com/plastics

Sterile aerosol barrier (ﬁltered) pipette tips[2] thermoﬁsher.com/

pipettetips

[1] For saliva only.

[2] Also available at fisherscientific.com.

Chapter 1 Product information

Required materials not supplied

6MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

Methods

Procedural guidelines

•The plates provided in this kit are single use plates only. Do not reuse the plates.

•Ensure plates are stored upright for 24 hours before opening.

•Perform all steps at room temperature (15–25°C) unless otherwise noted.

•Yellowing of the Binding and Wash I Solution is normal and will not impact buer performance.

Guidelines for saliva collection

•Saliva samples must be collected, transported, and stored using appropriate procedures and

conditions. Improper handling of samples decreases performance and sensitivity for detecting the

target sequences.

•Extraction of nucleic acid from clinical samples must be performed according to the speciﬁed

methods listed in this procedure. Other extraction approaches and preprocessing systems have not

been evaluated.

•Positive and negative test controls must be included to accurately interpret patient test results.

•Ensure that there was no eating, drinking, smoking, chewing tobacco, chewing gum, brushing

teeth, or use of mouthwash for at least 30 minutes before giving a saliva sample.

•At least 30 minutes before saliva collection, rinse the mouth with water by swishing water for 10

seconds and swallowing the water to rid mouth of debris.

•Use the passive drool technique to pool saliva in the mouth, then drool into a collection device.

•Ensure only saliva is collected by using the passive drool technique, with no coughing or collection

of phlegm.

•For saliva collection volume, follow the saliva collection device manufacturers instructions for use.

•For raw saliva, collect a minimum of 1 mL.

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 7

Guidelines for oropharyngeal and nasopharyngeal swabs

•Oropharyngeal and nasopharyngeal samples must be collected, transported, and stored using

appropriate procedures and conditions. Improper handling of samples decrease performance and

sensitivity of the assay to detect the target sequences.

•Extraction and ampliﬁcation of nucleic acid from clinical samples must be performed according

to the speciﬁed methods listed in this procedure. Other extraction approaches and preprocessing

systems have not been evaluated.

•Patient samples must be collected according to appropriate laboratory guidelines. Only synthetic

ﬁber swabs with thin plastic or wire shafts should be used. Other types of swabs may contain

substances inhibitory to molecular tests.

•Positive and negative test controls must be included to accurately interpret patient test results.

Before you begin

Before each use of the kit

•Gently mix reagents in vials before use. Avoid creating bubbles.

•Examine the plate containing the beads prior to opening. Ensure that the beads are at the bottom

of each well. Hold the plate by the narrow end and sharply swing the sealed plate in a downward

motion 5 times. Hold the plate on the opposite side and repeat.

•To remove seals from preﬁlled plates, place the preﬁlled plate squarely onto the benchtop, secure

the plate with one hand and grasp the seal at the lower left corner with the other hand. Using a

gentle but steady motion, peel the seal o of the plate diagonally toward the upper right corner

without jostling the contents.

•If required by your assay, designate one well of the Sample Plate for a negative extraction control.

Warnings and precautions

The MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit workﬂow should be performed by

qualiﬁed and trained sta to avoid the risk of erroneous results. Use separate areas for the preparation

of patient specimens and controls to prevent false positive results. Specimens and reagents must be

handled under a laminar airﬂow hood or biological safety cabinet.

•Specimens should always be treated as if infectious and/or biohazardous in accordance with safe

laboratory procedures.

•Follow necessary precautions when handling specimens. Use personal protective equipment (PPE)

consistent with current guidelines for the handling of potentially infectious specimens.

•Always use pipette tips with aerosol barriers. Tips that are used must be sterile and free from

DNases and RNases.

•Do not eat, drink, smoke, or apply cosmetic products in the work areas.

•Modiﬁcations to reagents, protocols, or instrumentation are not permitted, and are in violation of

the In Vitro Diagnostic Directive 98/79/EC.

Chapter 2 Methods

Before you begin

8MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

•Do not use the kit after the expiration date.

•Dispose of waste in compliance with the local regulations.

•Safety Data Sheets are available upon request.

•Reagents must be stored and handled as speciﬁed in “Contents and storage” on page 5.

Prepare samples for extraction

Prepare raw saliva samples

1. Upon receipt of samples for extraction, dilute the raw saliva sample 1:1 by adding an equal volume

of 1X PBS pH 7.4 (without calcium or magnesium) to the tube and vortex well at maximum speed

for 1 minute.

2. Let the diluted raw saliva samples sit and settle for at least 30 minutes at 20°C to 25°C.

Note: Gradually, 2 fractions will form. Do not disturb the layers.

3. (Optional) Centrifuge the diluted raw saliva sample at 1,500 x g (3,000 rpm) for 5 minutes to

separate the large debris.

4. Transfer 200 µL from the top fraction of the diluted raw saliva sample to the Sample Plate.

Note: Pipet slowly to avoid large debris and precipitants from the lower fraction.

Proceed to “Prepare the Sample Plate, then process samples on the instrument” on page 10.

Prepare preserved saliva samples

1. Upon receipt of samples for extraction, let the preserved saliva samples sit and settle for at least

30 minutes at 20°C to 25°C.

Note: In some cases, large debris may start to settle to the bottom. A clear separation may not

always be visible.

2. (Optional) Centrifuge the preserved saliva sample at 1,500 x g (3,000 rpm) for 5 minutes to

separate the large debris.

3. Transfer 200 µL from the top fraction of the preserved saliva sample to the Sample Plate.

Note: Pipet slowly to avoid large debris and precipitants from the lower fraction.

Proceed to “Prepare the Sample Plate, then process samples on the instrument” on page 10.

Prepare swab samples for extraction

1. Upon receipt of samples, vortex the sample.

2. Transfer 200 µL of transport media to the sample plate.

Proceed to “Prepare the Sample Plate, then process samples on the instrument” on page 10.

Chapter 2 Methods

Prepare samples for extraction 2

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 9

Perform viral nucleic acid isolation

Viral nucleic acid isolation is performed using the KingFisher™ Flex Magnetic Particle Processor with

96 Deep-Well Head and the MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit with a

specimen volume of 200 µL.

IMPORTANT! Do not attempt to process more than the maximum volume allowed for each sample

type. Yields and quality will be reduced.

Set up the instrument

1. Ensure that the instrument is set up with the proper magnetic head and the proper heat block, as

indicated in the following table.

Component Type

Magnetic head 96 deep-well magnetic head

Heat block 96 deep-well heat block

IMPORTANT! Failure to use the proper magnetic head and heat block results in lower yields.

2. Ensure that the proper program (MVPIIDX_Ready_200_Flex_v1) has been downloaded from the

product page and loaded onto the instrument.

Prepare the Sample Plate, then process samples on the instrument

1. Remove the Sample Plate and the vial of Proteinase K from the product box.

2. Remove the sealing ﬁlm of the Sample Plate, then add 5 µL of Proteinase K to each well.

Chapter 2 Methods

Perform viral nucleic acid isolation

10 MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

3. Conﬁrm the KingFisher™ Flex instrument deck positions with the following table.

Note: Place the pre-ﬁlled plate as per the protocol and check the deck position number. Deck

positions are also denoted on labels on the short and long ends of the plate.

Deck

position Preﬁlled plate Contents Preﬁlled volume

MagMAX™Virus Binding Buer PF

(Sample Plate)

(Cat. No. 100104740)

Binding Solution

+ Proteinase K

(+extraction control if

needed) + sample

Proteinase K: 5 μL

Sample: 200 μL

Binding Buer: 265 μL

MagMAX™ DNA/RNA Binding

Beads PF (Bead Plate)

(Cat. No. 100104744)

Magnetic Beads in

diluent buer

Magnetic beads: 10 μL

Diluent buer: 190 μL

MagMAX™ Virus Wash Buer I PF

(Wash I)

(Cat. No. 100104741)

Wash I Wash Buer I: 500 µL

MagMAX™ Virus Wash Buer II PF

(Wash II)

(Cat. No. 100104742)

Wash II (80% EtOH) Wash Buer II: 1,000 µL

MagMAX™ Virus Elution Buer PF

(Elution Plate)

(Cat. No. 100104743)

Elution Solution Elution Buer: 65 µL

4. Select the MVPIIDX_Ready_200_Flex_v1 program on the instrument.

5. Click Start. Peel o the sealing ﬁlm of each pre-ﬁlled deep-well plate, input the tip comb into the

Wash Buer II plate, then place each plate into the corresponding deck position.

6. After the program runs for ~25 minutes, immediately remove the Elution plate. The extraction is

now complete.

IMPORTANT! If not used for immediate processing, seal the plate containing the eluate to

prevent evaporation. Place the Elution Plate on ice for immediate use in real-time RT‑PCR.

Note: If bead carryover is observed, place the elution plate on a magnetic stand to pellet the

beads, then pipette the eluate to a new 96-well plate for use in real-time PCR. Review the real-time

PCR results to determine if re-extraction is required.

To ensure reliable performance of the KingFisher™ Flex Magnetic Particle Processor, perform

preventative maintenance as instructed by the manufacturer.

Chapter 2 Methods

Perform viral nucleic acid isolation 2

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 11

Troubleshooting

Observation Possible cause Recommended action

Low or inconsistent yield Plates were stored incorrectly. Store plates in an upright position at room

temperature. Examine the plate or row

containing the beads before removing the seal

for an indication of how to proceed.

For plates that have been inverted, store them

upright for at least 24 hours, then check that

the beads form a tight dark pellet in the center

of the bottom of the well before unsealing the

plate.

For plates without beads in that were not

stored correctly, ﬂick the plates in a fast

downward motion to ensure that the materials

are in the well and not on the seal before

unsealing.

For plates that were stored inverted, the

beads are dry and uneven in the bottom

of the wells and or not fully resuspended.

Flick the plate in a fast downward motion

to remove the ﬂuid from the seal, then

gently vortex to resuspend the beads before

unsealing. A gentle brief centrifugation can

be performed after resuspension but is not

always necessary.

After removing the seal, there

were some wet or dry beads

on the seal.

For wet beads, carefully pipet the liquid from

the seal back to its proper well.

For dry beads, resuspend the dry beads in

Nuclease-Free water, then carefully pipet the

liquid from the seal back to its proper well.

There were bubbles in the

wells.

Centrifuge the plates to remove the bubbles

before use.

The wells were blocked. Remove any seal covering well openings and

blocking tip comb access. Seal remnants on

the plate edge will not interfere with tip comb

access and do not have to be removed.

If the seal has delaminated and left a

transparent seal over the well then rotate plate

180 degrees and peel diagonally from the

corner that is now on the bottom left. If the

problem persists, call technical support.

12 MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

Observation Possible cause Recommended action

Low or inconsistent yield

(continued)

Heat block was installed

incorrectly.

Install the correct deep well heat block.

Sample input limits were

below or above recommended

amounts.

Consult user guide for recommended sample

input ranges.

Appendix A Troubleshooting

Perform viral nucleic acid isolation A

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 13

Safety

WARNING! GENERAL SAFETY. Using this product in a manner not speciﬁed in the user

documentation may result in personal injury or damage to the instrument or device. Ensure that

anyone using this product has received instructions in general safety practices for laboratories and

the safety information provided in this document.

·Before using an instrument or device, read and understand the safety information provided in the

user documentation provided by the manufacturer of the instrument or device.

·Before handling chemicals, read and understand all applicable Safety Data Sheets (SDSs) and use

appropriate personal protective equipment (gloves, gowns, eye protection, and so on). To obtain

SDSs, see the “Documentation and Support” section in this document.

Chemical safety

WARNING! GENERAL CHEMICAL HANDLING. To minimize hazards, ensure laboratory personnel

read and practice the general safety guidelines for chemical usage, storage, and waste provided

below. Consult the relevant SDS for speciﬁc precautions and instructions:

·Read and understand the Safety Data Sheets (SDSs) provided by the chemical manufacturer

before you store, handle, or work with any chemicals or hazardous materials. To obtain SDSs, see

the "Documentation and Support" section in this document.

·Minimize contact with chemicals. Wear appropriate personal protective equipment when handling

chemicals (for example, safety glasses, gloves, or protective clothing).

·Minimize the inhalation of chemicals. Do not leave chemical containers open. Use only with

sucient ventilation (for example, fume hood).

·Check regularly for chemical leaks or spills. If a leak or spill occurs, follow the manufacturer

cleanup procedures as recommended in the SDS.

·Handle chemical wastes in a fume hood.

·Ensure use of primary and secondary waste containers. (A primary waste container holds the

immediate waste. A secondary container contains spills or leaks from the primary container.

Both containers must be compatible with the waste material and meet federal, state, and local

requirements for container storage.)

·After emptying a waste container, seal it with the cap provided.

·Characterize (by analysis if needed) the waste generated by the particular applications, reagents,

and substrates used in your laboratory.

·Ensure that the waste is stored, transferred, transported, and disposed of according to all local,

state/provincial, and/or national regulations.

·IMPORTANT! Radioactive or biohazardous materials may require special handling, and disposal

limitations may apply.

14 MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

Biological hazard safety

WARNING! BIOHAZARD. Biological samples such as tissues, body ﬂuids, infectious agents,

and blood of humans and other animals have the potential to transmit infectious diseases.

Conduct all work in properly equipped facilities with the appropriate safety equipment (for example,

physical containment devices). Safety equipment can also include items for personal protection,

such as gloves, coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, or

goggles. Individuals should be trained according to applicable regulatory and company/ institution

requirements before working with potentially biohazardous materials. Follow all applicable local,

state/provincial, and/or national regulations. The following references provide general guidelines when

handling biological samples in laboratory environment.

·Laboratory biosafety manual, fourth edition. Geneva: World Health Organization; 2020 (Laboratory

biosafety manual, fourth edition and associated monographs)

www.who.int/publications/i/item/9789240011311

·U.S. Department of Health and Human Services, Biosafety in Microbiological and Biomedical

Laboratories (BMBL), 6th Edition, HHS Publication No. (CDC) 300859, Revised June 2020

https://www.cdc.gov/labs/pdf/CDC-BiosafetymicrobiologicalBiomedicalLaboratories-2020-

P.pdf

WARNING! Potential Biohazard. If you use the kit with the automated nucleic extraction workﬂow,

the surface of the KingFisher™ puriﬁcation system may be considered a biohazard. Use appropriate

decontamination methods when working with biohazards.

Appendix B Safety

Biological hazard safety B

MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide 15

Documentation and support

Customer and technical support

Visit thermoﬁsher.com/support for the latest service and support information.

•Worldwide contact telephone numbers

•Product support information

–Product FAQs

–Software, patches, and updates

–Training for many applications and instruments

•Order and web support

•Product documentation

–User guides, manuals, and protocols

–Certiﬁcates of Analysis

–Safety Data Sheets (SDSs; also known as MSDSs)

Note: For SDSs for reagents and chemicals from other manufacturers, contact the

manufacturer.

Limited product warranty

Life Technologies Corporation and/or its aliate(s) warrant their products as set forth in the Life

Technologies' General Terms and Conditions of Sale at www.thermoﬁsher.com/us/en/home/global/

terms-and-conditions.html. If you have any questions, please contact Life Technologies at india-

info@thermoﬁsher.com.

16 MagMAX™ Dx Preﬁlled Viral/Pathogen Nucleic Acid Isolation Kit User Guide

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Thermo Fisher Scientific MagMAX Dx Prefilled Viral/Pathogen Nucleic Acid Isolation Kit User guide

Thermo Fisher Scientific MagMAX Dx Prefilled Viral/Pathogen Nucleic Acid Isolation Kit User guide

Thermo Fisher Scientific MagMAX Dx Prefilled Viral/Pathogen Nucleic Acid Isolation Kit User guide

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