Sop

Standard Operating ProceduresFor the laboratories

South West District

Prepared by:-

Dr.Harpreet KaurPathologist SWD

Dated – 18th Nov’2010

Procedures:Procedures:

GENERAL GUIDELINES:

The treating doctor (consultant /medical officer) of the patient prescribes the

investigations to be performed in the patient’s OPD card.

The patients go to the sample collection area / LAB.

The laboratory technicians collect the patient’s sample in the designated

sample collection area.

The laboratory technician immediately labels the collected sample with the

patient’s name, OPD number, sample type and then the samples are stored in

respective laboratory repository.

Test results are entered into the system and reports are generated at the

reporting area of the laboratory.

S.O.P. FOR PHLEBOTOMY/ FINGER PRICK PROCEDURE Patient should be given basic information about venipuncture / finger

prick and must be made comfortable. He should be in a relaxed position

and tell him that he is going to get a small prick.

Check the details on the request form. (Check if patient is fasting or not

– for the required investigations).

Prepare all the vaccutainers / syringes / lancet as per test required.

Prepare the vein and apply the tourniquet DO NOT tie too hard. / prepare

the finger for finger prick.

Apply spirit using a spirit swab as disinfectant.

Allow it to dry.

Open the syringe pack if using the syringe or lancet for finger prick or

prepare the needle and the holder for using vaccutainer for collection.

For Finger prick - Hold the ring or middle finger firmly from the base of

first digit, to concentrate the blood flow, give a firm prick using the

lancet. Do not press the finger to increase or cause the blood flow; let

the blood flow on its own.

For Venepuncture - Touch the point of the needle to the skin and give a

prick along the direction of the vein taking precautions avoiding a double

puncture. Never draw blood too hastily, let the free flowing blood fill

the syringe / vaccutainer till the required amount of blood is

collected.

If using a syringe blood collected should be dispensed in appropriate

vaccutainer tubes.

First release the tourniquet and then slowly withdraw the needle from the

vein.

Press the puncture site gently with cotton and ask the pt. to press for 2-4

mins. to prevent bleeding.

The used needle must be destroyed in the needle destroyer / or discarded

in the appropriate sharps container or the used syringe should be cut and

disinfected in 1% sodium hypochlorite and discarded in red colored bag.

The cotton swab should be discarded in yellow colored bag.

Check the patient-if there is no bleeding and ask the patient to leave.

VACUTAINERS FOR BLOOD COLLECTION

Red Lavender Grey Blue

All Biochemistry

All Serology

Dengue Serology

Malaria Serology

All Hormones

All Markers

All Haematology

Hb

DLC

TLC

C. Haemogram

Peripheral Smear

Sugar PT

APTT

Fasting Samples: FBS, Lipid Profile, Uric Acid (Overnight fasting For 10-12

hrs.)

Random samples: All other

PREPARATION OF A BLOOD SMEAR:

Mix sample well, either by inversion or by mechanical rocker. Remove

stopper holding tube away from face. Using two wooden applicator

sticks rim the tube and check for fibrin clots.

Holding a 1 X 3 inch slide in your left hand by the short side, place a 2-3

mm drop of mixed whole blood about 1/4 inch from the right side of the

slide, utilizing the wooden applicator sticks held in the right hand.

Alternate Method: Leave slide on a flat surface. Or a same size drop of

blood from the finger prick is to be placed at one end of the slide.

Place the slide containing the drop of blood on a flat surface and hold

securely.

Grasp a second slide (spreader slide) in the right hand between thumb

and forefinger.

Place the spreader slide onto the lower slide in front of the blood drop,

and pull the slide back until it touches the drop.

Allow the blood to spread by capillary action almost to the edges of the

lower slide.

Push the spreader slide forward at approximately a 30 degree angle,

using a rapid, even motion. The weight of the spreader slide should be

the only weight applied. Do NOT press down. Perform this step

quickly. The drop of blood must be spread within seconds or the cell

distribution will be uneven.

Characteristics of a Good Smear

1) Thick at one end, thinning out to a smooth rounded feather edge.

2) Should occupy 2/3 of the total slide area.

3) Should not touch any edge of the slide.

4) Should be margin free, except for point of application.

SOP FOR STAINING TECHNIQUES

Leishman’s Staining

Propose: To examine a smear

Reagents: Leishman’s Stain

Distil Water

Specimen: Blood, Body fluid etc.

Equipment: Glass slides

Pipettes

Procedure:

1. Make a smear in a clean glass slide and air dry .

2. Cover the smear with Leishman’s Stain.

3. Wait for 2 min.

4. Add double volume of distilled / buffer water .

5. Wait for 8 – 10 min.

6. Wash in running tap water.

7. Dry and observe under oil immersion.

Interpretation: As per microscopic findings.

SOP FOR ESR:

Sample preparation

Sample must be drawn from a clean vein puncture over a maximum period of 30s

without any excessiye venous stasis. The blood should be diluted either at the time of

vain puncture (4 vol blood taken directly into 1 vol of sterile sodium citrate

anticoagulant- diluents ) or taken first into a primary anticoagulant (EDTA 1.5mg/ml)

followed by dilution in the sterile sodium citrate.

Custom made ESR tubes facilitate both the procedures mentioned.

Sample so prepared must be used immediately, paying attention to the following

external agents that might alter ESR in the pre—analysis phase.

If not used immediately, the sample must be stored at 4 C and has to be used within 8

hours after collection.

Manual

1. Insert the ESR pipette into the blood collection tube (black top vaccutainer).

2. Using continuous force, push the pipette down to the bottom of the tube. The

blood will automatically rise into the pipette and stop at the zero mark.

3. Place the test unit in the ESR rack .

4. At the end of 60 minutes, the numerical results are read in millimeters directly

from the imprinted scale on the pipette.

SEROLOGY:

Include – HbSAg, Malaria, Typhi rapid, Syphilis card test, HCV,

Perform the test according to the instructions given on the test kits.

Discard according to BMW guidelines.

GLUCOMETER :-

First, set out your glucometer, a test strip, a lancet and an alcohol swab.

Check the code on the strip matches with the glucometer.

Turn on the glucometer and place a test strip in the machine when the

machine is ready. Watch the indicator for placing the blood to the strip.

Make sure patients hand is dry and wipe the area you've selected with an

alcohol swab and wait until the alcohol evaporates.

Pierce the finger tip and obtain a drop of blood. The type of drop of

blood is determined by the type of strip you are using (some use a

"hanging drop" of blood versus a small drop for strips that draw blood in

with a capillary action).

Place the drop of blood on or at the side of the strip.

The glucometer will take a few moments to calculate the blood sugar

reading.

You may use the alcohol prep pad to blot the site where you drew the

blood if it is still bleeding.

Tips:

Make sure you keep batteries in stock that fit your glucometer.

Keep your glucometer and test strips in a clean, dry place.

SOP FOR URINE ANALYSIS

Check request is for UPT, Urine routine and microscopy.

Provide the patient with the container for sample collection.

Receive the sample from the patient

Label the sample.

UPT is performed there only (according to the kit sop) and the patient is

asked to wait ten minutes and report is given to the patient then itself after

entering the details along with the report in the master register.

Sample for Urine R&M is taken and labeled, entry made in the master

register.

URINE ROUTINE AND MICROSCOPY

Physical examination is done on the sample

Sample is poured in a labeled test tube

Using the uristix for sugar and protein or multistix for others chemical test

is performed and reported according to kits directions.

Urine is centrifuged at 1500 x rpm for 15 mins. Supernatant is discarded in

the sample container and a wet mount is prepared form the sediment.

Wet mount is observed under the microscope and reported.

HAEMATOLOGY ANALYZERS

Majority of the Hematology investigations are performed on 3 part automated

analyzer

Used for hematology parameters.

Follow the instructions given in the instruction manual for cell counters

Check the machine for reagents, sufficient quantity of the reagent should

be present.

In order to avoid any error the quantity of the reagent should be adequate.

Reagent should be left at the room temperature (15 to 30 degree C) for 24

hour before using.

After unpacking the reagent, care should be taken to prevent entry of dust,

dirt bacteria etc.

No dust should enter the instrument.

Inspect that there are no broken tubing and power cord is plugged in the

outlet.

Waste tank should be checked daily.

Check for the printer paper.

Procedure

Specified amount of sample corresponding to the amount of EDTA

anticoagulant should be collected from the vein.

Tubes up to 80 mm height should be used.

Sample Analysis:

Mix the sample sufficiently, preferably on the mixer.

Care should be taken while removing the tube cover to prevent blood

scatter.

Set the tube to sample probe position specified and aspirate the sample.

Do not wipe the probe clean..

MAINTANENCE

Daily:.

1. E-Z cleaning.

2. System shut down.

Weekly:

1. Entire system cleaning.

All the maintenance schedules are given in the manual of the cell counters.

Kindly follow instructions provided by the manufacturer.

STANDARD OPERATING PROCEDURES – BIOCHEMISTRY

Majority of the biochemistry investigations are performed on semi-automated

chemistry analyzer.

The work instructions to run a test are available in the kit inserts supplied along with

the kits. The complete information about specimen collection, volume, methodology,

storage and stability, reagents- working solutions, calibration, reference ranges,

analytical sensitivity etc. can be taken from the kit inserts.

Maintenance of the instrument

Daily maintenance

The instrument must be operated under the environmental requirement.

After turning on the power or the lamp wait 15 – 20 minutes before

starting the test .After turning off the lamp ,wait about 10 min for the lamp

to cool down before turning it on again

Make sure the measurement is free of bubbles.

Wash the flow cell after finishing each kind of test.

Fill the flow cell with D/w before shutdown.

Handle the dispose after waste according to acceptable laboratory local

state and national standard

Empty the waste bottle (in accordance with BMW guidelines)

System shut down at the end of the day.

Weekly maintenance

Calibrate Aspiration volume once a week,-

Check Zero drift.

Filter check

Monthly maintanence

2ml – 4ml clean with hypochloride.

All the maintenance schedules are given in the manual of the cell counters.

Kindly follow instructions provided by the manufacturer.

DO’S & DON’TS OF LABORATORY

♦All universal precautions to be observed in the Laboratory.

1. Gloves to be worn while handling specimens and hands should be washed

after removing gloves.

2. Laboratory coats are worn at all times in the laboratory.

3. Protective gear like mask must be worn while handling potentially infectious

specimens.

4. Decontaminate all surfaces and reusable devices after use with 1% Sodium

Hypochlorite.

5. Never leave a discarded tubes or infected material unattended or unlabelled.

6. Cleaning of reusable slides and tubes after disinfecting in 1% Sodium

Hypochlorite.

♦ Instructions for Housekeeping staff:

1. Glove to be worn while cleaning equipments.

2. No equipment to be switched off without prior instructions.

3. Only trained Housekeeping staff to handle the Laboratory equipments.

♦ All samples to be considered potentially Biohazardous and treated with 1%

Sodium Hypochlorite.

♦Proper waste disposal to be done in the specified color coded bags.

♦Daily maintenance of all instruments to be done.

SAFETY REGULATIONS / INSTRUCTIONS IN LAB

Follow the manufactures Instructions.

Do not touch chemicals with your hands.

Dispose off chemicals only by recommended procedure.

Wear gloves & lab coat while doing the tests.

Wash hands thoroughly after collection.

Avoid mouth pipetting.

During centrifugation always close the lid.

At the end of the day put all the reagents back on the shelf .

Clean the works station with 1% bleach solution.

Do not eat / drink in the lab / do not store eatables in the lab.

Adequate and consultant water supply should be provided for

immediate washing and cleaning.

General Instructions

Cleaning of the work area

The work area should be decontaminated and cleaned with 1% sodium

hypochlorite solution daily in the morning and as required.

The mops should be washed daily and sun dried.

Person doing cleaning should wear gloves.

Equipment Maintenance

Equipment maintenance as per the instructions in the manual.

All the laboratory equipments should be calibrated once daily before Starts

and as per instruction manual.

Daily cleaning and waste removal from the equipments should be done as

per the instruction guidelines.