Sample collection , handling and transportation

Muhammad IlyasPhD scholar (Biochemistry)

Gauze pads

Adhesive bandage

Tray for assembling blood collection tools

Rack for holding blood tubes

Durable marker for writing on laboratorysample

Step 1a : Assemble equipment for collecting blood:

Laboratory sample tubes for blood collection

Blood sampling systems (Needle and syringe system, vacuum extraction system with holder, winged butterfly system (vacuum extraction) or winged butterfly system

Tourniquet (single-use)

Skin antiseptic solution: 70% isopropyl alcohol

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Step 1: Before entering patient room, assemble all equipment Step 1c : Fill out patient documentation:Label blood collection tubes with date of collection, patient name, and his/her identifier number.Do NOT forget to fill out necessary laboratory form

Secondary ContainerFor the shipment of samples to the National Central Laboratory follow Sample Shipment packaging requirements (see document "How to safely ship Emerging and Dangerous Pathogen sample Tertiary Container

Step 1d : Assemble materials for packaging of samples:Plastic leak-proof packaging containerDisposable (paper) towelsCooler or cold box, if sample requires refrigeration

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Step 2: Put on all personal protective equipment (PPE)DO NOT ENTER THE PATIENT AREA IF YOU DO NOT HAVE ALL PROTECTIVE GEAR ON

Step 2a: Perform hand hygiene. Duration of the entire procedure: 40-60 sec.

Step 2c: Put on face protection:

Step 2b: Put on a gown.

Quick tips: If the patient has respiratory symptoms, wear amedical mask underneath the face shield.

Step 2d: Put on gloves (over gown cuffs).

Wet hands with water and enough soap to cover all hand surfaces.

Rub hands, palm topalm,

Right palm over left dorsum with interlaced fingers and vice versa,

Palm to palm withfingers interlaced

Back of fingers to opposing palms with fingers interlocked

Rotational rubbing of left thumb clasped in right palm and vice versa,

Rinse hands with water.

Dry hands thoroughly with single use towel.

Put on a medical mask and eye protection (e.g. eye visor/goggles)OR

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Step 3a: Prepare room.Put infectious waste bags and leak- proof and

puncture resistant sharps container into patient room and make sure they are ready for use.

Place all blood collection equipment in a place that is easy to access.

Step 3d. Apply a tourniquet around the arm.Tie approximately 4–5 finger widths

Step 3g: When using vacuum extraction system with holder, insert the blood collector tube into the holder.Avoid pushing the collector tube past the recessed line on the needle holder or you may release the vacuum.

Step 3b: Identify and prepare the patient.Introduce yourself to the patient and explain what you will do with the blood

sample and why.Make sure that this is the correct patient from whom you wish to take the

blood sample.

Step 3c: Select the site, preferably at the bend of the elbow.Palpate the area; locate a vein of good size that is visible, straight and clear.The vein should be visible without applying a tourniquet

Step 3e: Ask the patient to form a fist so that the veins are more prominent

Step 3f: Disinfect the area where you will put the needle.Use 70% isopropyl alcohol.Wait 30 seconds for the alcohol to dry.DO NOT touch the

Step 3h: Anchor the vein by holding the patient’s arm and placing a thumb BELOW the place where you want to place the needle.DO NOT touch the disinfected site.DO NOT place a finger over the vein to guide the needle.

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Step 3j: When blood starts to flow, ask patient to open his/her hand.

Step 3k: Once sufficient blood has been collected (minimum 5ml), release the tourniquet BEFORE withdrawing the needle

Step 3l: Withdraw the needle gently.Give the patient a clean gauze or drycotton wool ball to press gently on thesite.Ask the patient NOT to bend the arm

Step 3m: Remove blood collector tube from holder and put into rack.

Step 3n: Put needle into leak-proof and puncture resistant sharps container.

If the sharps container DOES NOT HAVE a needle remover:Put the needle and holder into a sharps container.Do not remove the needle from the holder.Do not reuse the needle.

If the sharps container HAS a needle remover:Remove the needle following instructions on the sharps container.Put the holder into the infectious waste bag for disinfection.

Step 3o: Stop the bleedingand clean the skin.Do not leave patient until bleeding

has stopped.Put an adhesive bandage on the

site, if necessary.Step 3p: Put items that dripblood or have body fluids onthem into the infectious

waste bag for destruction.

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Hospital waste can be broadly be defined into 2 categories

Risk Waste

• Infectious Waste

• Pathological Waste

• Sharps

• Pharmaceutical Waste

• Chemical Waste

• Radioactive Waste

Non – Risk Waste

Non – risk waste is that whichis comparable todomestic garbage presents nogreater

normaland

risk,therefore, than waste from

a normal home i.e.

• Paper• Packaging• Food Waste

Type of Waste

Treatment:• Incineration• Chemical Disinfection• Autoclaving• Encapsulation• Microwave irradiation etc.,

Final Disposal• Landfill• Burying inside Premises• Discharge into Sewer etc.,

Waste Disposal

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• Pressurized gas containers• Large amounts of reactive chemical waste• Radioactive waste• Silver salts or radiographic waste• Halogenated plastics (e.g. PVC)• Mercury or cadmium• Ampoules of heavy metals

Waste MinimizationWaste not to be Incinerated

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Colour code for Waste Segregation

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• Different color coding has to be assigned to variouswaste for effective segregation, as:

Non-Risk waste.Risk waste with Sharps. Risk Waste without sharps.

Black :Red:Blue:v.Yellow: Radioactive waste

Green: Chemicals like Mercury & Cadmium

• All this segregation should be done by the individual user.

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Step 4a: Take the blood tube from the tray and wipe the blood tube with a disposable paper towel.

Step 4c: Place all items that came into contact with blood into the infectious waste bag for destruction.

Step 4: Prepare blood sample for transport

Step 4d: Protect the sample from breaking during transport by wrapping the tube of blood in a paper towel.

Step 4e: Ask the designated assistant to approach the patient room, without entering.This person should have gloves on.This person should come close to you holding the open plastic leak-proof

packaging container.This person should not enter the patient room.

Step 4f: The person who has collected the blood sample should put the wrapped tube of blood into the plastic leak- proof packaging container.Be careful not to touch outside of leak-proof plastic tube with

gloves.

Step 4: Prepare blood sample for transport

Labeling specimens for transportation

Patient’s nameClinical specimenUnique ID number (Research/Outbreak)Specimen typeDate, time and place of collectionName/ initials of collector

Blood for smears

CollectionCapillary blood from finger prick

make smearfix with methanol or other fixative

Handling and transportTransport slides within 24 hoursDo not refrigerate (can alter cell morphology)

Blood for cultures

CollectionVenous bloodinfants: 0.5 – 2 ml children: 2 – 5 ml adults: 5 – 10 ml Requires aseptic techniqueCollect within 10 minutes of feverif suspect bacterial endocarditis: 3 sets of blood culture

Blood for cultures

Handling and Transport

Collect into bottles with infusion broth

change needle to inoculate the brothTransport upright with cushion

prevents hemolysisWrap tubes with absorbent cotton Travel at ambient temperatureStore at 4oC if can’t reach laboratory in 24h

Serum

CollectionVenous blood in sterile test tube

let clot for 30 minutes at ambient temperatureglass better than plastic

HandlingPlace at 4-8oC for clot retraction for at least 1-2 hoursCentrifuge at 1 500 RPM for 5-10 min

separates serum from the clot

Serum

Transport 4-8oC if transport lasts less than 10 daysFreeze at -20oC if storage for weeks or months before processing and shipment to reference laboratoryAvoid repeated freeze-thaw cycles

destroys IgM To avoid hemolysis: do not freeze unseparated blood

Cerebrospinal fluid (CSF)

Collection

Lumbar puncture Sterile tubesAseptic conditions Trained person

CSF

Handling and transportation

Bacteriapreferably in trans-isolate medium, pre-warmed to 25-37°C before inoculation

OR

transport at ambient temperature (relevant pathogens do not survive at low temperatures) Virusestransport at 4-8oC (if up to 48hrs or -70oC for longer duration)

Stool samples

Collection:Freshly passed stool samples

avoid specimens from a bed panUse sterile or clean container

do not clean with disinfectantDuring an outbreak - collect from 10-20 patients

Rectal swabs

Advantageconvenientadapted to small children, debilitated patients and other situations where voided stool sample not feasible

Drawbacksno macroscopic assessment possibleless material available not recommended for viruses

Stool samples for viruses

Timing

within 48 hours of onset

Sample amount

5-10 ml fresh stool from patients (and controls)

Methods

fresh stool unmixed with urine in clean, dry and sterile container

Storage

refrigerate at 4oC; do not freeze

store at -15oC - for Ag detection,polymerase chain reaction (PCR)

Transport

4oC (do not freeze); dry ice for (Ag detection and PCR)

Stool samples for bacteria

Timing

during active phase

Sample amount and size

fresh sample and two swabs from patients, controls and carriers (if indicated)

MethodCary-Blair medium For Ag detection/PCR – no transport medium

Storagerefrigerate at 4oC if testing within 48 hours, -70oC if longer; store at -15oC for Ag detection and PCR

Transport

4oC (do not freeze); dry ice for Ag, PCR detection

Stool samples for parasites

Timing

as soon as possible after onset

Sample amount and size

at least 3 x 5-10 ml fresh stool from patients and controls

Method

mix with 10% formalin or polyvinyl chloride, 3 parts stool to 1 part preservative

unpreserved samples for Ag detection and PCR

Storage

refrigerate at 4oC; store at -15oC for Ag detection and PCR

Transport

4oC (do not freeze); dry ice for antigen detection and PCR

Throat swab (posterior pharyngeal swab)

Hold tongue away with tongue depressorLocate areas of inflammation and exudate in posterior pharynx, tonsillar region of throat behind uvulaAvoid swabbing soft palate; do not touch tongueRub area back and forth with cotton or Dacron swab

WHO/CDS/EPR/ARO/2006.1

Nasopharyngeal swab

Tilt head backwardsInsert flexible fine-shafted polyester swab into nostril and back to nasopharynxLeave in place a few secondsWithdraw slowly; rotating motion

WHO/CDS/EPR/ARO/2006.1

Naso-pharyngeal aspirate

Tilt head slightly backwardInstill 1-1.5 ml of VTM /sterile normal saline into one nostrilUse aspiration trapInsert silicon catheter in nostril and aspirate the secretion gently by suction in each nostril

WHO/CDS/EPR/ARO/2006.1

Sputum

CollectionInstruct patient to take a deep breath and cough up

sputum directly into a wide-mouth sterile containeravoid saliva or postnasal discharge1 ml minimum volume

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Step 5a: Remove the gloves.

1.Grasp the outer edge of the 1st glove and peel it off.

2.Hold the 1st glove in 3. Remove 2nd glove fromthe gloved hand and drag the inside, creating aa bare finger under the "bag“ for both gloves and 2nd glove. throw it in waste bag for disposal.

Step 5b: Remove the gown

Untie the gown

Step 5c: Perform Hand hygiene. Duration of the entire procedure: 40-60 sec.

Step 5: Remove Personal Protective Equipment (PPE)

Step 5d: Take off face protectionRemove face shield from behind.If it is a reusable face shield, place it in an infectious waste bagfor disinfection.If it is a disposable face shield, place it in an infectious waste bag for destruction.Optional: If wearing a medical mask, remove the medical mask from behind, starting with the bottom strap, and place it in a infectious waste bag for destruction.

When wearing goggles and a mask:Remove goggles from behind.If reusable goggles, place it in an

infectious waste bag for disinfection.If disposable goggles, place it in an

infectious waste bag for destruction.Remove the medical mask from

behind, starting with the bottom strap, and place it in an infectious waste bag for destruction

Step 5e: Perform Hand hygiene. Duration of the entire procedure: 40-60 sec.

Quick Tips:Place all reusable equipment into a separate infectious waste bag for disinfection.

When collecting blood samples from multiple patients:Change gloves between each patient.Wash hands between each patient.DO NOT WASH GLOVED HANDS.DO NOT REUSE GLOVES.