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Collection and transport of clinical samples in laboratorries

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Sheshkant Poudyal
Sheshkant Poudyal

This document provides guidelines for proper collection, transport, and preservation of laboratory samples. It emphasizes that samples must be collected and handled aseptically to avoid contamination, and transported quickly to the laboratory in leak-proof containers using appropriate preservatives or transport media to maintain sample integrity. Accurate labeling and paperwork are also important. Adhering to these procedures helps ensure sample quality and accurate test results.

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Collection and transport of clinical samples in laboratorries
COLLECTION & TRANSPORT OF LABORATORY SAMPLES
AIM OF SAMPLE COLLECTION • Accuracy -To ensure that analytical results obtained are representative of actual analyte conc. in patient and, thus of his/her physiological/pathological state. • Safety • Minimize patient discomfort and complications • Avoid recollection
GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Should provide proper guidelines for collection of samples • All diagnostic information depends on quality of sample received • If sample collection, transport, media are not proper, it will give false results • Collect sample before administering antimicrobial agents when possible. • Collect sample with as little contamination from indigenous microbiota as possible to ensure that the sample will be representative of the infected site.
GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Utilize appropriate collection devices. Use sterile equipment and aseptic technique to collect specimens to prevent introduction of microorganisms during invasive procedures. • Clearly label the specimen container with the patient’s name and identification number. Always include date and time of collection and your initials. • Collect an adequate amount of specimen. Inadequate amounts of specimen may yield false-negative results. • Identify the specimen source and/or specific site correctly so that proper culture media will be selected during processing the laboratory
GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Collect specimens in sturdy, sterile, screw-cap, leak proof containers with lids that do not create an aerosol when opened. • Follow universal precaution guidelines. Treat all specimens as potentially biohazardous. • Laboratory workers should use appropriate barrier protection (such as gloves and laboratory coat or gown) when collecting or handling specimens. If splashing may occur, protective eyewear, face masks, and aprons may be necessary. • Minimize direct handling of specimens in transit from the patient to the laboratory. Use plastic sealable bags with a separate pouch for the laboratory requisition orders or transport carriers (for example, small buckets with rigid handles).
GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Do not contaminate the external surface of the collection container and/or its accompanying paperwork. • Label High risk Specimens eg: • Sputum with suspected Tuberculosis • Fecal samples suspected with Cholera, Typhoid • Serum when suspected with HIV/ HBV/HCV, infections
A REQUEST FORM SHOULD INCLUDE • Name ,Age, Sex, Address • CR/ Lab No/IPD/ OPD No ,Ward ( eg: ICU/CCU/FSW/ER) • Time and Date, • Urgent / Routine • Type of specimen • Investigation needed • History of patient • Provisional diagnosis • Doctor’s name and sign
EXAMPLES OF BIOLOGICAL SPECIMEN • Blood- venous/ arterial/ capillary- whole blood ; serum ; plasma • Urine Faeces • Saliva Solid tissues • Amniotic fluid Cerebrospinal fluid • Synovial fluid Peritoneal fluid • Pleural fluid Pericardial fluid
BLOOD SAMPLE COLLECTION •Take proper precaution (gloves) • Avoid contamination • Palpate vein • Apply disinfectant • Use sterile needle and syringe • Collect 5-10 ml blood for two sets of culture each • In children collect 2-5 ml • Infuse it into BHI after cleaning with spirit
SAMPLE REJECTION CRITERIA • Samples brought by unauthorized person • Sample not properly transported ( packaging, temperature, delayed transit) • Sample not labeled/label illegible • Label not marching with that of lab requisition form • Sample not accompanying of lab requisition form • Sample not accompanying proper/complete lab requisition form • Sample : insufficient quantity (to test/to repeat test/ back up)
SAMPLE REJECTION CRITERIA, contd… • Sample leakage • Haemolysed sample • Lipaemic sample • Sample turbid( contaminated) • Sample coming/collected beyond stipulated period • Sample coming beyond working hours(at odd hours) to the central lab • Sample collected in improper container
SPECIMEN TRANSPORT • Reliable, Rapid and Relevant Transportation • Within 2 hours of collection • Containers should be leak-proof • Separate section for paperwork • Special preservatives or holding media • Biohazard label
CONTAINERS FOR SAMPLE COLLECTION FOR FAECES: • Universal container • Spoon attached to the inside of the screw cap
FOR URINE: • Universal container for small quantities • For larger quantities 250 ml wide mouthed screw-capped bottles FOR SPUTUM: • No Universal container to be used • Squat ,wide-mouthed disposable containers should be used
FOR BLOOD: • Without anticoagulant for serological examination • With EDTA for parasitological Examination BLOOD CULTURE BOTTLE: • At least large enough to hold 50ml of liquid medium,which it is issued from laboratory, plus 5-10ml of patient’s blood.
FOR SEROUS FLUIDS: • Universal container • Addition of 0.3ml of 20% solution sodium citrate to the container prior to autoclaving (with the cap fitted) is recommended for collection of fluids that may coagulate on standing • This avoids difficulty in performing cell counts or centrifuging procedure with such fluids
SWABS: Swabs suitable for taking Specimens of exudates from the throat Nostril, ear , skin, wounds and other accessible lesions consist of a sterile pledget (eg:gauze) of absorbent material, usually cotton-wool or synthetic fiber, mounted on a thin wire of stick
SWABS FOR SPECIAL PURPOSE : • Baby swabs • Per-nasal swabs • Post-nasal swabs • Laryngeal swabs • High vaginal and cervical swabs • Serum coated cotton wool swab
EYE: • Conjunctival Swab • Corneal Scrapping • Anterior Chamber & Vitreous Culture EAR: • Inner Ear • Outer Ear UPPER RESPIRATORY TRACT: • Oral Swab Nasal Swab • Nasopharyngeal Swab Laryngeal Swab • Throat Swab
LOWER RESPIRATORY TRACT: • Sputum • Trans Tracheal Aspiration • Bronchio-alveolar lavage • Bronchial Brush GASTRIC LAVAGE / OTHER BODY FLUIDS: • Pleural Fluid • Peritoneal Fluid • Cerebro-Spinal Fluid • Pericardial Fluid • Synovial Fluid
Transport media for stool specimens • Cary-Blair All enteric organisms • Stuart All enteric organisms • Amies All enteric organisms • Alkaline peptone water Vibrios • V-R fluid Vibrios • Buffered glycerol saline All enteric organisms except Vibrios & Campylobacter
URINARY TRACT INFECTION(UTI): • Clean uro-genital area with soap and water, then rinse with water, make dry. FEMALES: • hold labia apart and begin voiding in commode, • after several ml have passed, collect midstream MALES: Clean glans area with soap and water, • then rinse with water, • retract foreskin; after several ml have passed, • collect midstream Urine
GENITAL TRACT SAMPLES CERVICAL SWAB (PAP SMEAR): FEMALE URETHRAL SWAB: HIGH VAGINAL SWAB: MALE URETHRAL SWAB: PROSTATE SWAB:
CYTOLOGY / BIOPSY / AUTOPSY SAMPLES • Histology In bottle containing a suitable fixative. Fixative volume should be more than 10 to 15 times of the specimen. • Cervical Samples / PAP Smears Dipped in a bottle containing a suitable fixative. Smeared slides should be proper labeled.
PRESERVATION OF SPECIMENS IN TRANSIT • The specimen must be properly treated both during its transport to the lab and from the time the serum has been separated until it is analyzed • For some tests, specimens must be kept at 4 degree Celsius until the specimens are analyzed
PRESERVATION OF SPECIMENS IN TRANSIT • Specimens for Ammonia, Blood gas analysis as pCO2, PO2 , Blood pH, Acid Phosphatase, Lactate, Pyruvate etc. are transported to the lab by placing the specimen container in ice water. • For thermally labile constituents: Serum/ plasma should be separated from cells in a refrigerated centrifuge
PRESERVATION OF SPECIMENS IN TRANSIT • Specimens for bilirubin & carotene must be protected from both the day light or fluorescent light • Plasma or serum should be separated as soon as possible (within 2 hours) • If it is impossible to centrifuge the specimen within 2 hour, the specimen should be held at room temperature rather than at 4 degree Celsius.
PRESERVATION OF SPECIMENS IN TRANSIT • If delay for analysis, the separated serum should be stored in capped tubes at 4 degree Celsius until analysis. • If specimen for analysis is unstable at 4 degree Celsius, serum should be held at -20 degree Celsius. • For serum LDH isoenzymes, store the sample at room temperature rather than at 4 degree Celsius.
TRANSPORT OF SPECIMENS • Specimen tubes should be centrifuged with stoppers in place • Polypropylene & polyethylene containers are suitable for specimen transport • Glass & polystyrene containers should be avoided. • Container must be leak-proof & should have teflon lined screw cap. • Material of both the container & stopper must be inert.
TRANSPORT OF SPECIMENS • Solid carbon dioxide (dry ice) is the most convenient refrigerant material for keeping specimens frozen & temperature as low as - 70 degree Celsius. • One piece of solid dry ice (3”x4”x1”) in a container with 1” Styrofoam walls & a volume of 125 cubic inches will maintain a single specimen frozen for 48 hrs.
TRANSPORT VIA POSTAL SERVICE • The sending of specimens by post is governed by regulations laid down by the Postmaster General. • The specimen must be sent by letter post only and must be labeled clearly: “PATHOLOGICAL SAMPLE – HANDLE WITH CARE” . • The specimen must be in a sealed container packed in a wood or metal box which contains sufficient absorbent material such as sawdust or cotton wool to prevent any possible leakage should the container be damaged en route.
Collection and transport of clinical samples in laboratorries

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Collection and transport of clinical samples in laboratorries

  • 3. AIM OF SAMPLE COLLECTION • Accuracy -To ensure that analytical results obtained are representative of actual analyte conc. in patient and, thus of his/her physiological/pathological state. • Safety • Minimize patient discomfort and complications • Avoid recollection
  • 4. GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Should provide proper guidelines for collection of samples • All diagnostic information depends on quality of sample received • If sample collection, transport, media are not proper, it will give false results • Collect sample before administering antimicrobial agents when possible. • Collect sample with as little contamination from indigenous microbiota as possible to ensure that the sample will be representative of the infected site.
  • 5. GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Utilize appropriate collection devices. Use sterile equipment and aseptic technique to collect specimens to prevent introduction of microorganisms during invasive procedures. • Clearly label the specimen container with the patient’s name and identification number. Always include date and time of collection and your initials. • Collect an adequate amount of specimen. Inadequate amounts of specimen may yield false-negative results. • Identify the specimen source and/or specific site correctly so that proper culture media will be selected during processing the laboratory
  • 6. GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Collect specimens in sturdy, sterile, screw-cap, leak proof containers with lids that do not create an aerosol when opened. • Follow universal precaution guidelines. Treat all specimens as potentially biohazardous. • Laboratory workers should use appropriate barrier protection (such as gloves and laboratory coat or gown) when collecting or handling specimens. If splashing may occur, protective eyewear, face masks, and aprons may be necessary. • Minimize direct handling of specimens in transit from the patient to the laboratory. Use plastic sealable bags with a separate pouch for the laboratory requisition orders or transport carriers (for example, small buckets with rigid handles).
  • 7. GENERAL CONSIDERATION FOR PROPER SAMPLE COLLECTION • Do not contaminate the external surface of the collection container and/or its accompanying paperwork. • Label High risk Specimens eg: • Sputum with suspected Tuberculosis • Fecal samples suspected with Cholera, Typhoid • Serum when suspected with HIV/ HBV/HCV, infections
  • 8. A REQUEST FORM SHOULD INCLUDE • Name ,Age, Sex, Address • CR/ Lab No/IPD/ OPD No ,Ward ( eg: ICU/CCU/FSW/ER) • Time and Date, • Urgent / Routine • Type of specimen • Investigation needed • History of patient • Provisional diagnosis • Doctor’s name and sign
  • 9. EXAMPLES OF BIOLOGICAL SPECIMEN • Blood- venous/ arterial/ capillary- whole blood ; serum ; plasma • Urine Faeces • Saliva Solid tissues • Amniotic fluid Cerebrospinal fluid • Synovial fluid Peritoneal fluid • Pleural fluid Pericardial fluid
  • 10. BLOOD SAMPLE COLLECTION •Take proper precaution (gloves) • Avoid contamination • Palpate vein • Apply disinfectant • Use sterile needle and syringe • Collect 5-10 ml blood for two sets of culture each • In children collect 2-5 ml • Infuse it into BHI after cleaning with spirit
  • 11. SAMPLE REJECTION CRITERIA • Samples brought by unauthorized person • Sample not properly transported ( packaging, temperature, delayed transit) • Sample not labeled/label illegible • Label not marching with that of lab requisition form • Sample not accompanying of lab requisition form • Sample not accompanying proper/complete lab requisition form • Sample : insufficient quantity (to test/to repeat test/ back up)
  • 12. SAMPLE REJECTION CRITERIA, contd… • Sample leakage • Haemolysed sample • Lipaemic sample • Sample turbid( contaminated) • Sample coming/collected beyond stipulated period • Sample coming beyond working hours(at odd hours) to the central lab • Sample collected in improper container
  • 13. SPECIMEN TRANSPORT • Reliable, Rapid and Relevant Transportation • Within 2 hours of collection • Containers should be leak-proof • Separate section for paperwork • Special preservatives or holding media • Biohazard label
  • 14. CONTAINERS FOR SAMPLE COLLECTION FOR FAECES: • Universal container • Spoon attached to the inside of the screw cap
  • 15. FOR URINE: • Universal container for small quantities • For larger quantities 250 ml wide mouthed screw-capped bottles FOR SPUTUM: • No Universal container to be used • Squat ,wide-mouthed disposable containers should be used
  • 16. FOR BLOOD: • Without anticoagulant for serological examination • With EDTA for parasitological Examination BLOOD CULTURE BOTTLE: • At least large enough to hold 50ml of liquid medium,which it is issued from laboratory, plus 5-10ml of patient’s blood.
  • 17. FOR SEROUS FLUIDS: • Universal container • Addition of 0.3ml of 20% solution sodium citrate to the container prior to autoclaving (with the cap fitted) is recommended for collection of fluids that may coagulate on standing • This avoids difficulty in performing cell counts or centrifuging procedure with such fluids
  • 18. SWABS: Swabs suitable for taking Specimens of exudates from the throat Nostril, ear , skin, wounds and other accessible lesions consist of a sterile pledget (eg:gauze) of absorbent material, usually cotton-wool or synthetic fiber, mounted on a thin wire of stick
  • 19. SWABS FOR SPECIAL PURPOSE : • Baby swabs • Per-nasal swabs • Post-nasal swabs • Laryngeal swabs • High vaginal and cervical swabs • Serum coated cotton wool swab
  • 20. EYE: • Conjunctival Swab • Corneal Scrapping • Anterior Chamber & Vitreous Culture EAR: • Inner Ear • Outer Ear UPPER RESPIRATORY TRACT: • Oral Swab Nasal Swab • Nasopharyngeal Swab Laryngeal Swab • Throat Swab
  • 21. LOWER RESPIRATORY TRACT: • Sputum • Trans Tracheal Aspiration • Bronchio-alveolar lavage • Bronchial Brush GASTRIC LAVAGE / OTHER BODY FLUIDS: • Pleural Fluid • Peritoneal Fluid • Cerebro-Spinal Fluid • Pericardial Fluid • Synovial Fluid
  • 22. Transport media for stool specimens • Cary-Blair All enteric organisms • Stuart All enteric organisms • Amies All enteric organisms • Alkaline peptone water Vibrios • V-R fluid Vibrios • Buffered glycerol saline All enteric organisms except Vibrios & Campylobacter
  • 23. URINARY TRACT INFECTION(UTI): • Clean uro-genital area with soap and water, then rinse with water, make dry. FEMALES: • hold labia apart and begin voiding in commode, • after several ml have passed, collect midstream MALES: Clean glans area with soap and water, • then rinse with water, • retract foreskin; after several ml have passed, • collect midstream Urine
  • 24. GENITAL TRACT SAMPLES CERVICAL SWAB (PAP SMEAR): FEMALE URETHRAL SWAB: HIGH VAGINAL SWAB: MALE URETHRAL SWAB: PROSTATE SWAB:
  • 25. CYTOLOGY / BIOPSY / AUTOPSY SAMPLES • Histology In bottle containing a suitable fixative. Fixative volume should be more than 10 to 15 times of the specimen. • Cervical Samples / PAP Smears Dipped in a bottle containing a suitable fixative. Smeared slides should be proper labeled.
  • 26. PRESERVATION OF SPECIMENS IN TRANSIT • The specimen must be properly treated both during its transport to the lab and from the time the serum has been separated until it is analyzed • For some tests, specimens must be kept at 4 degree Celsius until the specimens are analyzed
  • 27. PRESERVATION OF SPECIMENS IN TRANSIT • Specimens for Ammonia, Blood gas analysis as pCO2, PO2 , Blood pH, Acid Phosphatase, Lactate, Pyruvate etc. are transported to the lab by placing the specimen container in ice water. • For thermally labile constituents: Serum/ plasma should be separated from cells in a refrigerated centrifuge
  • 28. PRESERVATION OF SPECIMENS IN TRANSIT • Specimens for bilirubin & carotene must be protected from both the day light or fluorescent light • Plasma or serum should be separated as soon as possible (within 2 hours) • If it is impossible to centrifuge the specimen within 2 hour, the specimen should be held at room temperature rather than at 4 degree Celsius.
  • 29. PRESERVATION OF SPECIMENS IN TRANSIT • If delay for analysis, the separated serum should be stored in capped tubes at 4 degree Celsius until analysis. • If specimen for analysis is unstable at 4 degree Celsius, serum should be held at -20 degree Celsius. • For serum LDH isoenzymes, store the sample at room temperature rather than at 4 degree Celsius.
  • 30. TRANSPORT OF SPECIMENS • Specimen tubes should be centrifuged with stoppers in place • Polypropylene & polyethylene containers are suitable for specimen transport • Glass & polystyrene containers should be avoided. • Container must be leak-proof & should have teflon lined screw cap. • Material of both the container & stopper must be inert.
  • 31. TRANSPORT OF SPECIMENS • Solid carbon dioxide (dry ice) is the most convenient refrigerant material for keeping specimens frozen & temperature as low as - 70 degree Celsius. • One piece of solid dry ice (3”x4”x1”) in a container with 1” Styrofoam walls & a volume of 125 cubic inches will maintain a single specimen frozen for 48 hrs.
  • 32. TRANSPORT VIA POSTAL SERVICE • The sending of specimens by post is governed by regulations laid down by the Postmaster General. • The specimen must be sent by letter post only and must be labeled clearly: “PATHOLOGICAL SAMPLE – HANDLE WITH CARE” . • The specimen must be in a sealed container packed in a wood or metal box which contains sufficient absorbent material such as sawdust or cotton wool to prevent any possible leakage should the container be damaged en route.