GUIDELINES FOR COLLECTION, PACKAGING AND TRANSPORT OF SPECIMENS FOR

TESTING FOR HIGH RISK VIRAL PATHOGENS

ICMR-NATIONAL INSTITUTE OF VIROLOGY
PUNE 411021, MAHARASHTRA
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1. Purpose: This document describes the processes for collection, packaging and transport of clinical
specimens to Maximum Containment (Biosafety Level 4) laboratory of the ICMR-National Institute
of Virology (NIV), Pune for diagnosis of high risk viral pathogens affecting humans.

2. Scope: This document is intended for use by clinicians/laboratories planning to collect, pack and
transport clinical specimens from patients suspected to have high risk viral pathogens.

3. Responsibilities:
• The clinician, based on the clinical presentation of the patient and associated
epidemiological factors, shall decide the necessity for laboratory testing and the type(s) of
specimen to be collected for high risk pathogen infection.
• A physician, trained nurse or medical laboratory technologist would be responsible for
collecting the right type of specimen as per standard protocols and test requirements.
• A trained medical laboratory technical staff or another staff member supervised by a clinical
microbiologist/scientist/laboratory supervisor shall perform the standard triple packaging of
the specimens and make arrangements for their safe and prompt shipment to the testing
laboratory.
• All documentation related to the sample collection, packing and transportation should be
checked and signed by the laboratory supervisor and the head of the laboratory.

4. Laboratories authorized for testing for high risk viral pathogens in India: The Ministry of Health &
Family Welfare, Govt. of India has designated the following entity as the apex laboratory for testing for
high risk viral pathogens affecting humans, in India:

Primary contact person:

a. Dr. Pragya D. Yadav, Scientist-‘F’ & Group Leader,

Maximum Containment Facility,
ICMR-NIV, Microbial Containment Complex,
130/1, Sus Road, Pashan, Pune 411021
Tel: Office: 020-26006111;
Email id: [email protected]

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 Secondary contact person:

a. Dr. Rima R. Sahay, Scientist ‘C’,

Maximum Containment Facility,
ICMR-NIV, Microbial Containment Complex,
130/1, Sus Road, Pashan, Pune-411021.
Tel: Office: 020-26006160;
Email id: [email protected]

5. Routing of samples
All samples intended for laboratory testing for high risk viral pathogens should be forwarded
through the head of the department/institute and the State Surveillance Officer.

6. Preparations for sample collection

• Prior to collecting the clinical specimen, the technologist should verify the identity of the
patient and the type and volume of sample to be collected.
• The individual collecting the clinical specimen must wear a full set of recommended
personal protective equipment (PPE) including a laboratory gown, two pairs of gloves,
goggles, N95 mask, and booties.
• All necessary instruments and supplies required for sample collection should be assembled
prior to starting the work. These include spirit swabs, tourniquet, and syringes with
needles/vacutainers/butterfly needles, serum collection tubes/gel separator tubes, tube
racks, labels, sample storage boxes, sterile swabs with viral transport medium, screw-
capped vials/urine collection cups, etc.
• Screw-capped, sturdy, leak-proof, polypropylene tubes of reliable quality and preferably
with an O-ring cap, should be used to collect the samples.
• The used syringes and needles should be discarded into a sharps container, and precautions
must be taken against needle-stick injuries.
• The sample vials should carry clear labels specifying the patient’s name, age, gender,
hospital ID and date and time of sample collection.

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7. Types of samples to be collected
• Blood, throat swab, urine and cerebrospinal fluid (CSF) samples should be collected from the
affected individuals. Where available, autopsy tissue specimens should also be forwarded.
• At least 5 mL of whole blood should be collected in a plain tube.
• Throat swabs should be collected and transported in Viral Transport Medium (VTM) in a
sterile, screw-capped polypropylene tube.
• About 10-20 ml of urine samples should be collected in to a sterile, screw-capped, leak-
proof and sturdy container.
• About 2-3 ml of CSF specimens should be collected by lumbar puncture into a sterile tube,
and The CSF specimen should not be frozen.
• Samples for PCR testing should preferably be sent in dry ice, or at -20°C, using hard-frozen
gel packs.
• Tissue biopsy specimens, if available, should be transported in a screw-capped, leak-proof
container with a small volume of Viral Transport Medium, preferably in dry ice or at -20°C.
• Where feasible, the blood, throat swab and urine samples of the patient contacts/caregivers
should also be forwarded for testing.

8. Packing of specimens
a. Sample has to be packed following the principles of the standard triple packaging system.
b. Primary container: Screw-capped vials to be used for sending serum/CSF. Vials should be labeled
with the sample number and test required. Keep the vials in upright position.
c. Secondary Container: A leak-proof plastic container (e.g., zip-lock bag, 50 mL centrifuge tube or
other similar containers) can be used to place the primary vials. Plastic pouches, if used, should be
heat-sealed or tightly closed by applying rubber bands. There should be enough absorbent
material (paper napkins/old newspaper) packed around the vials to absorb all fluid in case of
breakage or leakage. More than one vial can be placed in secondary container.
d. Outer Container: If a plastic pouch was used as a secondary container, it can now be placed inside
a separate plastic container and adhesive tape should be applied around the lid of the container.
This container can be placed in another plastic pouch and sealed with rubber bands before placing
it in the outer container. A Thermocol box or vaccine carrier containing ice/gel packs could be used
as the outer container. [The smallest overall external dimension of the outer container shall be 10
x 10 x 10 cm.]
e. Before despatching the outer container, Bleach can be used for surface disinfection. A 1:100
dilution of bleach or 5% Lysol solution should be used to clean the outer surfaces of the container.
f. The complete Case Report Form carrying all requisite information should be placed inside a sealed
plastic cover on the outer container.
g. Label the outer container as follows:
• The sender’s name, address and telephone number
• The receiver’s name, address and telephone number
• “BIOLOGICAL SUBSTANCE, CATEGORY B”
• Whom to contact in an emergency, with telephone number

9. Documents required:
• Patients history sheet to be part of document
• To be prepared and signed by the sender: A packing list/proforma invoice that includes the
sender's and the receiver’s address, the number of packages, details of contents, weight,
value
• To be prepared by the sender or the shipper’s agent: An air waybill for air transport or
weight, value equivalent documents for road, rail and sea journeys.

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 Sample packing and transport is to be done as per WHO Guidance on regulations for the Transport of
Infectious Substances 2013–2014.

A diagrammatic representation of the Standard Triple Packaging is given below:

Before dispatching the samples, the outer container may be wiped using 1:100 diluted bleach or 5%
Lysol solution.

Upon completion of sample packing, the personnel should perform thorough handwashing.

10. Shipment of samples

 The samples should be shipped only after prior intimation to the referral laboratory.
 The ICMR-NIV can assist in sample shipment from difficult locations, and the laboratories can
contact the above persons.

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 ASSEMBLING THE ITEMS REQUIRED FOR CLINICAL SAMPLE
PACKAGING AND TRANSPORT
1. Appropriate sample vials 2. Absorbent material and other accessories 1)

3. A leak-proof, sturdy, secondary container (e.g., ziplock

pouch, cryobox, 50 mL centrifuge tube, plastic 4. Hard-frozen Gel Packs
container)

5. A suitable outer container (e.g., thermocol box, ice-box, hard-board box)

(minimum dimensions: 10 x 10 x 10 cm)

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 INSTRUCTIONS ON SAMPLE PACKAGING AND TRANSPORT
STEP 1: ARRANGING THE SAMPLE VIALS

A B C

Wear full set of Personal Protective Seal the neck of the sample vials using Cover the sample vials using
Equipment and identify the labelled parafilm to prevent leakage during absorbent material to contain leakage,
sample tubes. transit. in the event of a breakage.

STEP 2: ARRANGING THE PRIMARY VIALS WITHIN A STURDY, LEAK-PROOF SECONDARY CONTAINER

A B

Option 1: Using a cryo-box as a secondary container. (Seal Option 2: Using a 50-mL centrifuge tube as a secondary
the lid of the box after arranging the samples, using cello.) container. (Seal the neck of the tube using cello.)
C D

Placing the centrifuge tube inside a zip-lock pouch Placing the zip-lock pouch inside a sturdy plastic container.
(Seal the neck of the container using cello.)

[Note: Sample vials can also be placed inside a zip-lock pouch, covered in absorbent material and secured by
heat-sealing or rubber bands. Then, the zip-lock pouch should be placed inside another plastic pouch and
secured.]
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STEP 3: ARRANGING THE OUTER CONTAINER

A B C

Option 1: Option 2: Placing the completed Case Report

Form/Request Form inside a leak-
Using a thermocol box as an outer Using a hard-board box as an outer
proof, zip-lock pouch
container and placing the container and placing the
secondary container within it, secondary container and the gel
surrounded by hard-frozen gel packs
packs
D E
Documents to accompany:
1) Packing list/Proforma Invoice
2) Air way bill (for air transport)
(to be prepared by sender or
shipper)
3) Value equivalence document
(for road/rail/sea transport)

Securing the zip-lock pouch with Attaching the labels:

the Case Report Form on the outer  Sender’s Address and contact
container number;
 Consignee’s Address and contact
number;
 Emergency Contact’s name and
number;
 ‘Biological Substance-Category B’;
 ‘UN 3373’; Orientation Label;

Note:
1. A vaccine-carrier/ice-box can also be used as an outer container
2. The minimum dimensions of the outer container should be 10 x 10 x 10 cm (length x width x height)

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