The document outlines protocols for the collection and preservation of viral specimens, emphasizing the importance of timing, site, technique, volume, and proper labeling. It details various specimen types required for diagnosing viral infections, including blood, swabs, and cerebrospinal fluid, along with specific storage conditions and preservation methods like viral transport media (VTM). Additionally, it stresses the necessity of accurate documentation and safety measures for handling potentially infectious samples.

1. VIRAL SPECIMEN COLLECTION AND
PRESERVATION
Mr. Manoj Mahato

2. Specimen collection for viral Disease
 At the correct time (stage of infection)
 From the correct site (site of infection)
 In the correct way
 In the adequate volume (for all tests needed)
 In the proper containers (sterile and chemically clean)
 Correctly labelled (name, date, type of specimen) and with
(age, sex, epidemiological data – vaccinations, etc.)

3. Transport specimen should be
 Protected from breaking
 Protected from light
 At adequate temperature:
-48 hrs at +4°C (refrigerator, wet ice)
-more than 48 hrs at -70°C (dry ice)
Note : must not be frozen at -20°C

4. Viral Transport Media (VTM)
 VTM contains: -buffer (adequate pH)
-saline (adequate ion concentration)
- proteins (albumin or gelatin)
- antibiotics and fungicides
 VTM is used to: Preserve viral infectivity within the specimen
Prevent specimen from drying Stop the growth of bacteria
and fungi
* MEM, Hank’s solution, Stuart’s * EDTA, Sodium citrate, heparin
for (blood)

5. Common types of
Specimen
 Swab
 Cerebrospinal fluid
 Nasopharyngeal secretions
 Faeces
 Blood
 Tissue Biopsy
 Skin and ulcer specimens

6. SWAB
 Pernasal swab e required to diagnose respiratory
syncytial virus infection, and when necessary to investigate
infections caused by influenza and parainfluenza viruses,
measles, rubella virus, adenoviruses, and enteroviruses
 Preserved in VTM

7. Cerebrospinal fluid
 Collect ml of fluid in a dry, sterile, leak-proof container.
 Refrigerated immediately at 4 OC.
 Transport in an insolated cold box.
 Cerebrospinal fluid is required to investigate some
arbovirus infections.
 When meninges are infected, the CSF will contain
lymphocytes and the CSF total protein will be raised

8. Skin and ulcer specimens
 Collect skin scrapings in a dry sterile container.
 Refrigerate immediately at 4OC,
 Transport in a cold box.
 If measles or rubella is suspected, the virus is more
likely to be isolated from a per nasal swab.
 Skin specimens are required to investigate infections
caused by herpes simplex viruses, rabies viruses

9. Blood For Serological Tests
 Collect the sample within 5 days of the onset of the symptoms
 Second sample 5-10 days later.
 Blood sample is required for investigating newborn infant with
congenital defects, determining immunity to rubella or other
viruses.
 Blood sample is required when testing for hepatitis B surface
Antigen (HBsAg) and HIV-1 and HIV-2.
 After the blood collected, collect in leak-prove container,
refrigerate at 4C
 Transport it in ice box to the virology lab.

10. Anticogulated Blood For Culture
 Collect 7-10 ml of blood into a sterile tube or bottle
which contains heparin, gently mix the blood with the
anticoagulant.
 Viruses which can be cultured from blood include dengue
viruses and some arboviruses.

11. Faeces
 Place about 4-8 g of faeces (small spoon) in a clean, dry, leak-
proof container (not a waxed carton)
 Deliver to the laboratory as soon as possible.
 If there is likely to be a one gram of faeces in 9 ml of
phosphate buffered saline.
 Label and keep at -20 OC.
 Send to the virology laboratory in a cold box
 Faeces specimens are required to diagnose viralgastroenteritis
specially that caused by rotaviruses, and to investigate
poliomyelitis and other diseases caused by enteroviruses

12. Nasopharyngeal secretions
 Collect a specimen by passing a sterile cotton wool swab,
attached to a pent piece of thin wire, through the floor
of the nasal cavity as far as the nasopharynx.
 After few seconds, slowly withdraw the swab and
immerse the swab in a container of sterile VTM, cutting
off swab wire to allow the bottle cap to be replaced
tightly
 Used for the isolation of influenza viruses, measles virus,
and respiratory syncytial viruses.

13. Tissue Biopsy
 Autopsy Tissue samples collection: Tissue Biopsy or
Autopsy
 1.Sterile instruments should be used.
 2.Collect 3 x 5 mm and placed in tube with VTM
 Storage In the refrigerator (2 – 6.0° C) for up to one
week.
 Can be stored frozen (≤ -70.0 ° C) indefinitely.

14.  Specimens such as urine and sputum are generally
unsuitable for viruses' isolation.
 All samples which may contain dangerous and highly
infectious viruses must be labeled HIGH RISK with a
red marker pen, and handled with great care.

15. Labeling of specimens:
 This should be including:
 Patient's name and number.
 Hospital or health centre.
 Type of specimen.
 Date and time of collection.
 Container tops should be sealed with adhesive tape
to prevent slacking during transit.

16. Information to specimens
 A request form should include the fallowing information:
 1. Name, place, age, number of the patient, and details of
any recent travel.
 2. Type of specimen, if two, mention as the first of second.
 3. Investigation required.
 4. Full clinical information: nature duration, and severity of
disease.
 5. Details about immunization and antimicrobial therapy.
 6. Name and address of the hospital or health centre.

17. THANK YOU