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            SAMPLE COLLECTION MANUAL OF LABORATORY SERVICES
                    SIR T HOSPITAL, BHAVNAGAR 

Issue No: 04

Issue Date: 01/11/2019

Copy No: 01

Holder’s Name: Laboratory Director

CONTROLLED COPY


II. CONTACT INFORMATION

Postal Address: Sir T General Hospital, Jail Road, Bhavnagar, Gujarat Pin-364001

Routine Day Enquiries

Contact details of the sections are:

SectionsContact NoExtension
Biochemistry+91-278-2423501162/1163
Microbiology+91-278-2423501035
Pathology+91-278-2423501072
Histopathology+91-278-2511511267

                        IV  RELEASE AUTHORIZATION

The sample collection Manual is prepared as per ISO 15189:2012. This is the property of Sir T Hospital & Government Medical College Bhavnagar and issued under the authority of the undersigned.

Laboratory Director Issue No:
Laboratory Services Issue Date:
Sir T Hospital Copy No:
Bhavnagar CONTROLLED COPY

                               V. AMENDMENT   RECORD   SHEET
Sr NoPage NumberDate of AmendmentAmendment detailReason of AmendmentAmendment Approved by
1
2
3
4

                               VI. DISTRIBUTION LIST

The following are the authorized holders of the controlled soft copy only of sample collection manual.

CONTROLLED COPY NO.NAME/DESIGNATION OF THE HOLDER OF CONTROLLED SOFT COPY
Copy No.1LSSTH Director
Copy No.2Technical Manager
Copy No.3Quality Manager
Copy No.4Pathology Laboratory
Copy No.5Microbiology Laboratory
Copy No.6Biochemistry Laboratory
Copy No.7Sample collection center
                                VII  TABLE OF CONTENTS
QSP NoContentsPage No
1Title Page
2Contact information
3Introduction
4Release authorization
5Amendment record sheet
6Distribution list
7List of abbreviations
8Table of contents
9Documentary Procedure of Patient identification
10Documentary Procedure of Venipuncture
11Documentary Procedure of Capillary blood collection
12Documentary Procedure of Arterial Blood collection
13Documentary Procedure of Urine Specimen collection
14Documentary Procedure of Safety and Infection Control
15Documentary Procedure of Specimen Handling & Transport
16Sample acceptance & rejection criteria

Purpose: Laboratory testing provides information about a patient's health to assist Physicians in diagnostic and therapeutic decisions. Specimen Integrity is dependent on accurate pre-analytical processes to include patient preparation, specimen collection, handling, and transportation. Improper collection and handling of samples can give erroneous results and compromise the care of the patient. These guidelines cover some of the key steps in handling blood samples to provide optimal specimens for testing.

Principle: There are multiple Pre-Analytical factors associated with the handling and processing of laboratory specimens that can lead to test result inaccuracy. Strict adherence to all phases of proper collection and processing is essential for accurate test results.

Pre-Analytical Factors:

1. Specimen Collection:

2. Specimen Handling/Processing:

3. Specimen Transportation


                       VIII Documentary Procedure of Patient identification

Purpose: To ensure accurate patient identification before phlebotomy to prevent mislabeling of specimens and ensure patient safety.

Responsibility: Phlebotomy personnel

Procedure: The phlebotomist's role requires a Greet the patient and identify yourself and indicate the procedure that will take place. Prior to any patient procedure, proper identification is a priority. Proper identification is a three-step process.


                          Documentary Procedure of Patient Preparation

                           Documentary Procedure of Venipuncture
                           

Collection of blood from the vein is called venipuncture. Also referred to as phlebotomy. Assemble the necessary materials and equipment:

GlovesVacutainer tube
Vacutainer tube holder Two-way needle
Routine: 21 gaugeChildren: 23 gauge
Sharp containerTourniquet
gauze pads or cotton70% alcohol or suitable skin antiseptic
Sterile syringe and needle (if the syringe method is used)

Wear the Personal Protective Equipments:


Selection of Site of Puncture:

  1. Medial Cubital- First choice. Anchored and easy to penetrate
  2. Cephalic - On the outside surface, Well anchored
  3. Basilic - Not well anchored, tends to roll, painful

Preparation of Site:

Venous Occlusion:

Procedure for collection using Evacuated system:


Order Of Draw: Specific order for Blood collection tubes is followed to avoid cross-contamination of additives between tubes. The recommended order of draw is:

  1. Plain (Red top)
  2. SST (red-gray or gold top). Contains a gel separator and clot activator.
  3. Sodium heparin (dark green top)
  4. PST (light green top). Contains lithium heparin anticoagulant and a gel separator.
  5. EDTA (lavender top)
  6. ACDA or ACDB (pale yellow top). Contains acid citrate dextrose.
  7. Oxalate/fluoride (light gray top)

NOTE: Tubes with additives must be thoroughly mixed, otherwise erroneous test results may be obtained when the blood is not thoroughly mixed with the additive.


                Venepuncture technique using needle and syringe
  1. The needle is sharp
  2. The syringe moves smoothly
  3. There is no air left in the barrel

Venepuncture in Children A syringe should be the tuberculin type or should have a 3- mL capacity, except when a large volume of blood is required for analysis


                   Documentary  Procedure of Capillary blood collection

Also referred to as micro blood samples collection/dermal puncture

Indications:

Sites of Puncture:


Materials Required: Cotton or gauze pads 70% alcohol or other skin antiseptic Sterile disposable lancet (Automatic lancet) Blood collection tubes

Technique:


                      Factors  interfere with test results

Hemolysis can interfere with many test results.

Indwelling Lines or Catheters:

Hemoconcentration: An increased concentration of larger molecules and formed elements in the blood may be due to several factors:

Prolonged Tourniquet Application:


                  Documentary Procedure of Arterial Blood collection  

Arterial Blood collection

Choice of site:

Equipment and supplies:

Procedure for arterial blood sampling using radial artery:

Caution and Care during arterial blood sampling:

Sampling errors:

* Presence of air in the sample * Collection of venous rather than arterial blood * An improper quantity of heparin in the syringe, or improper mixing after blood is drawn * A delay in specimen transportation.


Documentary Procedure of Urine Specimen collection

Random: The normal composition of urine varies considerably during a 24-hour period. Most reference values are based on analysis of the first urine voided in the morning. This specimen is preferred because it has a more uniform volume and concentration, and its lower pH helps preserve the formed elements. Submit a first morning specimen whenever possible. Urine for pregnancy testing should be a first morning voiding, or a random specimen with a specific gravity of at least 1.010. Note the time of collection of the specimen on the test request form and on the label of the container. Submit urine for pregnancy testing in a plastic vial with no preservative. To reduce contamination, the specimen submitted for urinalysis should be a clean catch “midstream sample.” Urinalysis specimens must be submitted in a yellow capped urine transport tube containing preservative.

24-Hour Urine Collection: Proper collection and preservation of 24-hour urine specimens is essential for accurate test results. Patients are carefully instructed in the correct procedure.

A collection container and detailed instructions are given to the patient. If a urine preservative is required, it is ensured that the designated preservative is added in the urine collection container at the start of the collection. The patient is instructed that the preservative may be toxic and caustic and that it should not be spilled or discarded. Record any medication that the patient is receiving on the test request form. Mix the urine well, measure the volume of the 24-hour collection and record volume on both the test request form and the transport vial. Transfer the required volume into a plain urine transport vial. Add any additional required preservative and mix well. Do not send the entire urine collection.

Note: For those analyses requiring the addition of 6N HCI or other preservatives, add the preservative at the start of collection. Have the patient collect each specimen in a smaller container and carefully pour the urine into the 24-hour container to avoid any possible acid burns to the patient. Be sure to mix urine thoroughly before removing the aliquot.

Instructions to the Patient:

  1. Avoid alcoholic beverages and vitamins for at least 24 hours before starting to collect urine, and during the collection period. Do not discontinue medications unless instructed to do so by your physician. Inform the laboratory which medications you are taking.
  2. Do not exceed your normal intake of liquids or change your dietary habits during the day before and the day of your collection unless your physician gives you specific instructions to do so.
  3. Empty bladder (void) into the toilet on the morning of the collection day. Do not include the first urine specimen of the day.
  4. Collect all subsequent urine voided for the next 24 hours and add to the container provided by the laboratory. The last sample collected should be the first specimen of the following morning at the same time as the previous morning's first voiding.
  5. Keep the urine in a cool place. Refrigerate if possible.
  6. Deliver to the referring healthcare provider promptly.
  7. Preservative may have been added to this container. This may be caustic. Please be careful not to spill any of the preservative.

—-

                Documentary Procedure of Safety and Infection Control                                   

* Because of contacts with sick patients and their specimens, it is important to follow safety and infection control procedures.

Self Protection: Standard Precautions:

  1. Wear gloves and a lab coat or gown when handling blood/body fluids.
  2. Change gloves after each patient or when contaminated.
  3. Wash hands frequently.

If you prick yourself with a contaminated needle:

Follow the institution's guidelines regarding treatment & follow-up. Note: The use of prophylactic zidovudine following blood exposure to HIV has shown effectiveness (about 79%) in preventing seroconversion.

Patient Precaution:


                 Documentary Procedure of Specimen Handling & Transport 

Quality laboratory results begin with proper collection and handling of the specimen submitted for analysis. Correct patient preparation, specimen collection, specimen packaging and transportation are of vital importance. Specific specimen requirements for each test are provided. Submit the quantity specified for each test requested. Tests will be canceled as “QNS” (Quantity Not Sufficient) when the sample volume is inadequate and will be indicated as such on the report. As a general rule, the volume of blood drawn should equal 2-1/2 times the amount of serum plasma required. For example, to obtain 4mL serum or plasma, draw at least 10mL blood. When inappropriate or insufficient specimens are submitted, the laboratory will store them and contact the requesting facility to request that the specimen be recollected. Documentation of this action will appear on the report. Specimen Storage: Instructions for storage and transport of specimens for individual tests are listed. All serological samples are stored for 72 hrs and all positive serum samples are stored for 15 days then discarded, all HIV positive serum samples are stored for 3 months as per the NACO guidelines. Maintain the specimen at the temperature indicated for each test until specimens are delivered to the laboratory. Most specimens require refrigeration unless otherwise indicated. Most of the specimens require refrigeration unless otherwise indicated. All other samples are retained for 24 hrs and then kept in hypochlorite for 6 hrs before discarding.

Specimen Requirements Blood and Bone Marrow: * Collect whole blood specimens in EDTA, Heparin or ACD vacutainer tubes. Do not dilute bone marrow aspirate specimens with peripheral blood. * Maintain at room temperature. Do not refrigerate. * Deliver specimens to the laboratory within 24 hours. Obtain and transport Monday

Stool Specimen Collection (24, 48 or 72-Hour)

Specimen Requirements Blood and Bone Marrow:

Blood sample acceptance criteria

  1. Properly filled Requisition forms.
  2. Properly labeled samples.
  3. Details on Requisition form should be matched with sample container details
  4. Proper Vacutainers
  5. Appropriate quantity of sample.
  6. ABG sample & Plasma sample should not be clotted
  7. Spill over the sample should not be there.
  8. Sample will be received within 4 hours of collection.

Urine sample acceptance criteria

  1. Properly filled Requisition forms.
  2. Properly labeled samples.
  3. Details on Requisition form should be matched with sample container details
  4. Appropriate quantity of sample.
  5. Sample containers should not be open, they should be tightly packed.
  6. Sample spillage should not be there.

Blood sample rejection criteria

  1. Improperly filled Requisition forms.
  2. Improperly labeled samples.
  3. Details on Requisition form is not matched with sample container details
  4. Improper Vacutainers
  5. Insufficient sample volume / Overfilled sample 6. Clotted sample (Whole blood/Plasma)
  6. Spill over sample.
  7. Hemolyzed sample.
  8. Sample received after 4 hours of collection.
  9. Diluted Sample

Urine sample rejection criteria

  1. Improperly filled Requisition forms.
  2. Improperly labeled samples.
  3. Details on Requisition form is not matched with sample container details
  4. Insufficient sample volume.
  5. Improper sample container
  6. Spill over sample.

Sample acceptance Exceptions

A.Insufficient sample volume

B.Hemolyzed sample

C.If blood is collected in EDTA Vacutainer: Bilirubin, Cholesterol, Triglyceride, ALT, AST, Creatinine, Urea can be performed. However it is unsuitable for Coagulation study,Enzyme assay, Hormonal assay & electrolyte analysis or methods that require electrolyte or metallic cofactors.


         Standard Operating Procedure Of Specimen Handling And  Transport 

Quality laboratory results begin with proper collection and handling of the specimen submitted for analysis. Correct patient preparation, specimen collection, specimen packaging and transportation are of vital importance.

Specific specimen requirements for each test are provided. Submit the quantity specified for each test requested. Tests will be cancelled as “QNS” (Quantity Not Sufficient) when the sample volume is inadequate and will be indicated as such on the report. As a general rule, the volume of blood drawn should equal 2-1/2 times the amount of serum plasma required. For example, to obtain 4mL serum or plasma, draw at least 10mL blood. When inappropriate or insufficient specimens are submitted, the laboratory will store them and contact the requesting facility to request that the specimen be recollected. Documentation of this action will appear on the report. Specimen Storage: Instructions for storage and transport of specimens for individual tests are listed. All serological samples are stored for 72 hrs and all positive serum samples are stored for 15 days then discarded, all HIV positive serum samples are stored for 3 months as per the NACO guidelines. Maintain the specimen at the temperature indicated for each test until specimens are delivered to the laboratory. Most specimens require refrigeration unless otherwise indicated. Most of the specimens require refrigeration unless otherwise indicated. All other samples are retained for 24 hrs and then kept in hypochlorite for 6 hrs before discarding.

Stool Specimen Collection (24, 48 or 72-Hour) Special gallon-size containers are available upon request for the collection of stool specimens. Refer to specimen requirements for each analyte.

Specimen Requirements Blood and Bone Marrow:

STANDARD OPERATING PROCEDURES (LSSTH/SCM/SOP/SHT)2/3

Provide the following information on the Test Request Form

  1. Patient name, sex and age
  2. Patient Social Security
  3. Identification number
  4. Collection time
  5. Collection date
  6. Sample origin or location
  7. Ordering physician

HISTOPATHOLOGY SAMPLE COLLECTION AND TRANSPORTATION

  1. 10% Formalin must be taken from Histopathology Laboratory or prepared in Operation theatre by the following formula: Method for preparation of 10%formalin:

40%formaldehyde:100ml + Tap water:900ml

  1. Immediately after removal of organ/biopsy, it should be put in a wide mouthed screw-capped specimen jars of plastic containing 10%formalin fixative.
  2. Volume of 10%formalin should be 10 times the volume of tissue.
  3. The histopathology requisition form should be complete with following details name of the patient, age, sex, requisition no., ward no., name of the incharge Doctor, history or clinical diagnosis, nature of the specimen, date of operation.
  4. Specimens are sent to the Central Pathology Laboratory-transported by hospital class IV staff or patients only in case of OPD patients, between 9a.m.-5p.m.in a capped bottle of suitable size in formalin.
  5. On receipt of specimen/biopsy, details are verified by the technical assistant and if necessary, returned for correction.
  6. If all details are okay, then details are entered in receiving register and a card is given to OPD patients except skin and ENT patients indicating date and time for collection of report form histopathology laboratory.
  7. Specimens are transported to histopathology laboratory in a closed container along with forms by class IV and on-duty driver at 12 noon and 4.30pm.
  8. On arrival to histopathology laboratory, specimens and forms are again cross checked by laboratory assistant or technician.
  9. If all details are okay, then specimen is opened, verified, the formalin changed and transported in new bottle by technical assistant/technician if a. Foamy, b. Reddish, c. Top up with formalin if not 10 times the volume of the tissue, d .If organ is too large for container or, e. If container is cracked or not closing properly.
  10. Receipt details are entered in the Histopathology receiving register.

Storage of Histopathology specimens: All specimens of histopathology are routinely stored for 3 months and afterwards they are discarded.

CYTOPATHOLOGY SAMPLE COLLECTION AND TRANSPORTATION

FluidPap
Collection TimingRoutine8:00 am to 4:00 pm Transfer within 1 hourDuring OPD timing
Saturday8:00am to 12:00 pm. Transfer within 1 hour
Sunday& HolidaysAvoid collection.If required to collect, aliquot and keep in ward refrigerator at 2-8 C and transport at 9:00 am on next working day
AmountIdeal is 10 ml directly from tapping line. Minimum 2 ml. If taken from receptacle, aliquot should be mixed well1 slide endocervix 1 slide ectocervix
ContainerClosed cap bottle. BAL fluid in BAL containerEither in wide mouthed closed bottle or wrapped in paper
TransportTo be sent to laboratory within 1 hour of collection Ward Ayah or servant will bring to laboratory along with requisitionPatient will bring bottle with slides or dried slides (wrapped) to cytology laboratory
Receiving*R1 will receive it. Check the demographics and sign the register.*R1 will receive the slides, check requisition form and give pap number on case paper.
ResponsibilityCollection Clinical Resident/Consultant Clinical Resident/Consultant
Transport Class IV of ward Class IV of OPD/Patient’ relative
Receiving R1 Pathology R1 Pathology

7.1 STANDARD OPERATING PROCEDURE OF RECEIVING ROUTINE SPECIMENS IN HISTOPATHOLOGY

  1. if foamy.
  2. Reddish.
  3. Top up with formalin if not 10times the volume of the tissue.
  4. If organ is too large for container or if container is cracked or not closing properly.
               ''STANDARD OPERATING PROCEDURES (LSSTH/SCM/SOP/FSCP)1/1''

7.2 STANDARD OPERATING PROCEDURE OF FNAC SAMPLE COLLECTION PROCEDURE

            ''STANDARD OPERATING PROCEDURES (LSSTH/SCM/SOP/SRC)1/5''

7.3 SAMPLE REJECTION CRITERIA BLOOD SAMPLE REJECTION CRITERIA

CRITERIA FOR SAMPLE REJECTION IN HISTOPATHOLOGY

  1. Patient’s Name , Reg. no, Ward
  2. Specimen name

Incomplete requisition form

  1. Patient’s Name, Reg. no, Ward/Unit
  2. Name of unit Incharge
  3. Related history and investigation
  4. Clinical diagnosis
  5. Specimen name
  6. Name and signature of authorized person

FNAC REJECTION CRITERIA

       ''STANDARD OPERATING PROCEDURES (LSSTH/SCM/SOP/SRC)2/5''

REJECTION CRITERIA OF MICROBIOLOGY SPECIMENS

Purpose: To provide Microbiology specimen rejection criteria & the procedure to follow when rejecting specimens for process.

General Principle: Proper collection & transport of specimens is critical to the quality of results produced by the microbiology Laboratory & cannot be overemphasized. The validity of all diagnostic information produced in the lab is contingent on the quality of the specimen received. Consequences of poorly collected and/or poorly transported specimens include failure to isolate the causative organism, and recovery of contaminants or normal flora could lead to improper treatment of the patient. The saying is “garbage in—garbage out”.

General Consideration: When a specimen is received in the laboratory, it is subjected to scrutiny & must meet certain minimum requirements before being accepted for testing. A specimen shall not be rejected until appropriate efforts have been made to correct the problem, but it may be rejected for failing to meet acceptability criteria. If, needed the specimen is rejected, it is listed on the microbiology disposition log for rejected specimens, along with reason for rejection & documentation of notified personnel.

Rejection criteria: A. SPECIMEN RECEIVED WITHOUT A LABEL OR WITH AN IMPROPER LABEL

B.SPECIMEN IMPROPERLY COLLECTED OR TRANSPORTED

C.SPECIMEN CONTAINER EXHIBITS GROSS EXTERNAL CONTAMINATION OR LEAKING SPECIMEN

D.SPECIMEN NOT OF SUFFICIENT QUANTITY:If specimen has multiple orders, request priority listing of tests.

E.SPECIMEN MUST BE RECEIVED IN A TIMELY MANNER Order recollection if specimen has exceeded recommended limits for transport.

Sr. No.Specimen typeTestRejection CriteriaReport Comment
1StoolC&S1.Not submitted in enteric pathogen transport media. 2.Multiple specimens collected from the same in-patient the same day(only one specimen per patient per test per day is to be processed).3.All formed stools except when S. typhi requested.1.Specimen not received in enteric transported medium.2.Multiple specimens received. Only the most recently collected specimen has been processed.3.Formed stool received. Test cancelled.
2SputumC&SMore than 25 squamous epithelial cells/lower power fieldGreater than 25 squamous epithelial cells per low power field.
3UrineC&S * Condom catheter * Foley catheter tips & bags * Leaking specimens * Inappropriate/swabs/non sterile container * > 24 hr delay before specimen received * Duplicate specimens (more than one processed urine within 24 hrs)“Specimen unsuitable for culture…. Because of….add qualifier.”
4Dry swabsC&SIf received in the lab > 1 hr after collection.“Specimen not received in transport medium & was in transit for > 1 hr.”
5Blood cultures, tissues, Sterile body fluids e.g. CSFC&SIf received in the lab > 48 hr after collection. Process specimen. EnterReport Comment:”Specimen was received in the lab>48 hrs after it has been collected; results should be interpreted accordingly.”
6All specimen type except: Tissue Sterile, body fluid,Brain & liver abscess, Aspirated pus, Bone & soft tissue biopsies with diagnosis of gas gangrene, necrotizing fasciitis or necrotizing cellulitis Anaerobic culture Not submitted in special anaerobic transport media. “No Anaerobic swab received; anaerobic culture not done.”
7Blood For serology* Sample hemolysed.* Not submitted in Red vacuette * So kindly repeat the sample.” * Please collect blood in Red vacuette.
Name of Laboratory : Laboratory Services Sir T. Hospital (LSSTH),Bhavnagar
Document Name: Documented procedure for primary sample collection & handling(Sample collection manual)
Unique ID: LSSTH/B/Central/DP/7.2.4/22
Issue No. : 01Issue Date :30/04/2024Copy No. :01
Authorized by:Laboratory director Review Date:23/09/2024Reviewed by:Deputy Quality Manager