Sample collection and safety procedure in laboratory

Sample collection and safety procedure in laboratory

1. WELCOME 1

2. Presented by: Dr. Priyanka Buragohain Guided by : Dr Swapan Kr Chakraborty . Prof. & HOD Dr Anup Baishya Associate Prof. Dr. Hemen Kalita Assistant Prof. Dr. Mrinal Baishya Lecturer SAMPLE COLLECTION AND SAFETY PROCEDURE IN LABORATORY 2

3. Why is sample collection necessary ? To study different investigative parameters 3

4. Blood Urine Stool Sputum Semen Cerebrospinal fluid Body fluids Gastric washings Ear/Eye/Nose/ Pernasal /Wound/ Throat/ Vaginal swabs Nasopharyngeal aspirate Fungal samples of hair, nail & skin etc . Biopsy material SAMPLES COLLECTED IN LAB 4

5. Haematological study : Blood R/E, Hb %, CBC count,PBS study, coagulation profile study, genetic study etc. Biochemical study : Blood sugar, LFT, Bl. Urea, Sr. creatinine , Sr. uric acid etc. Serology : CRP, VDRL, HIV, RA factor, ASO titre, widal test, etc. Bacteriological culture Immunological study : detection of antibodies and antigens, etc. Blood transfusion services : ABO grouping, cross matching of blood, etc. STUDY OF DIFFERENT INVESTIGATION FROM A SAMPLE OF BLOOD 5

6. For fasting BS- empty stomach with a preferable fasting of 8-10 hours For PPBS- a full meal or 75 gm of glucose intake For lipid profile- a fasting sample of not less than 12 hours, no fatty foods in previous meal No alcohol before sample collection PATIENT PREPARATION 6

7. BLOOD COLLECTION PROCEDURE 7

8. Veins : Anticubital vein, veins of arm, dorsum of hands, long saphenous vein, femoral vein, umbilical and scalp vein. Capillaries : Finger tips, lobule of ear, heel and thumb. Arteries : femoral artery . SITES FOR COLLECTION OF BLOOD 8

9. VENOUS BLOOD COLLECTION Make the patient sit Arrange the required equipments Examine the site of collection Locate the site Apply the torniquet Locate the vein Cleanse the area with an alcohol wipe Dry with gauze Feel and fix the vein by pressing down on the vein Remove the needle shield Approach the vein in the same direction the vein is running with the needle 15* to the participant’s arm Push the needle with bevel facing up Make sure that the needle is in the vein Loosen the torniquet and allow the needle to fill as much as needed Withdraw the needle out of the arm, press gauze firmly on the puncture Collect the blood in approprite container. Discard the needle into a sharps container. 9

10. Wipe the tip of the finger, using alcohol swab and let it dry. 3 rd and 4 th fingers of non dominant hands are preffered . Using a sterile lancet, make a skin puncture just of the centre of the finger pad. The puncture should be made perpendicular to the ridges of the fingerprint. Wipe out the first drop of blood. Free flowing blood may be collected by gently massaging the finger. Hold a small gauze pad over the puncture site for a couple of minutes to stop bleeding. In infant, gently rub the heel to warm it. Clean it and prick on the medial/lateral part of plantar surface to collect blood. CAPILLARY BLOOD COLLECTION 10

11. For blood gas analysis ARTERIAL BLOOD COLLECTION 11

12. Red top ADDITIVE None MOA blood clots and the serum is separated by centrifugation USES Chemistries, Immunology and serology, blood bank (cross matching) Gold top ADDITIVE None MOA SST contains a gel at the bottom to separate blood from serum on centrifugation. USES Chemistries, Immunology and serology ANTICOAGULANTS 12

13. Light green top ADDITIVE Lithium heparin MOA Anticoagulates with heparin. Plasma is separated with PST gel at the bottom of tube USES Chemistries Purple top ADDITIVE EDTA MOA Forms calcium salts to remove calcium USES Haematology and blood bank 13

14. Light blue top ADDITIVE Sodium citrate MOA Forms calcium salts to remove calcium USES Coagulation tests Green top ADDITIVE Sodium heparin or Lithium heparin MOA Inactivates thrombin and thromboplastin USES For lithium level, ammonia level 14

15. Dark blue top ADDITIVE EDTA MOA Tube is designed to contain no contaminating metals USES Trace element testing, toxicology Light grey top ADDITIVE Sodium fluoride and potassium oxalate MOA Antiglycotic agent preserves glucose upto 5 days USES Glucoses 15

16. Yellow top ADDITIVE ACD (acid citrate dextrose) MOA Complement inactivation USES HLA tissue typing, paternity testing, DNA studies Yellow black top ADDITIVE Broth mixture MOA Preserves viability of microorganisms USES microbiology- aerobes, anaerobes and fungi 16

17. Black top ADDITIVE Sodium citrate MOA Forms calcium salts to remove calcium USES ESR Orange top ADDITIVE Thrombin MOA Quickly clots blood USES STAT serum chemistries 17

18. Light brown top ADDITIVE Sodium heparin MOA Inactivates thrombin and thromboplastin contains no lead USES Serum lead determination Pink top ADDITIVE Potassium EDTA MOA Forms calcium salts USES Immunohaematology 18

19. White top ADDITIVE Potassium EDTA MOA Forms calcium salts USES Molecular/ PCR and bDNA testing 19

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21. Adult draw: M in 2 ml 10 ml red top &10 ml lavender top for antibody 16-20 ml for culturing Paediatric draw : 0.5 – 5ml for culturing Newborn : capillary SPECIMEN VOLUME 21

22. Wash hands thoroughly with soap and water. Dry completely Let first stream of urine to drain into toilet. Place a sterile container under the stream and fill the container. Do not touch the rim or inner surface of the container. Place and tighten lid on the container. Level and sent to laboratory. URINE SAMPLE COLLECTION 22

23. Prevent contamination Send urine within 1 hour for accurate culture result Can refrigerate for upto 24 hours if delay. Bagged = BAD, highly unreliable Voided clean catch= 80% - 90% accurate if perineum well cleaned and caught midstream. Catheterized = Most accurate and reliable. Supra pubic aspiration = very very accurate 23

24. Urine specimens for culture should be collected in C&S preservative tubes. Fill the tube upto the minimum fill line. Mix tube 8-10 times immediately after filling. Morning specimen is preffered . Maintain normal daily fluid intake, avoid alcohol. In case of 24 hr urine collection do not void on the collection container. In case of creatinine clearance 1 st hydrate the pt. By administering a minimum of 600 ml of water before the collection. Avoid coffee, tea and drugs 24

25. Position sample collection paper across the rim of toilet bowl. Make a bowel movement onto the collection paper . Avoid mixing with urine or water from toilet . Poke onto stool at six different sites . Collect in a neat, clean, wide mouthed jar. Do not clump, scoop, or fill the tube. Screw it tightly and level it. Store between 2-8 degree or room temperature 25

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27. Transport within 1 hour of collection or refrigerate upto 24 hours. Warm stools are best for detecting ova and parasites. Not recommended on patients who have been hospitalisd for >3 days & not admitted with a diagnosis of gastroenteritis. Specimen should be collected before antibiotic therapy is initiated. Diarrhoeal stool will always give good results. 27

28. Mouth should be pernished before sample collection. For fungal culture – 3 consecutively collected early morning specimens are recommended. For AFB – Collect 3 early morning specimens from a deep cough or 3 consecutively collected specimens, each collectad in 8-24 hour intervals with at least one being an early morning specimen. Sample should be collected in a sterile disposable, impermeable container. SPUTUM COLLECTION 28

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31. In Adult : L3-L4 level In small children : L4-L5 level or lower. Morning is preferred rather than late afternoon or evening. Always use stylette inside the needle. CEREBROSPINAL FLUID COLLECTION 31

32. Collected from large joints like knee, ankle, hip, elbow, wrist, and shoulder. 10- 20 ml flud to be obtained in 3 or 4 sterile tubes. Plain tube –gross examination, viscisity , mucin clot test EDTA tube- cell counts and microscopic study. Heparinised tube – microbiologic study Fluoride tube - glucose SYNOVIAL FLUID COLLECTION 32

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34. Do not remove more than 1 lt of fluid at a time. Collect in 3 EDTA tubes. PLEURAL FLUID COLLECTION 34

35. Collect in 3 tubes EDTA- gross and microscopic examination Plain- microbiologic examination Heparinised- chemical examination. Should be done under CT scan guidance. PERICARDIAL FLUID COLLECTION 35

36. Urinary bladder to be emptied Patient to lie in supine or semi reclined position. Large bore I.V. needle is introduced in the midway between symphysis pubis and umbillicus . Needle is connected with a rubber tubing hich drains the fluid into the container. 20-50 ml fluid is sufficient for diagnostic procedures. PERITONEAL FLUID COLLECTION 36

37. Lie the patient in supine position. Determine the position of foetus and pocket of amniotic fluid with ultrasound. Prepare the area (mother’s abdomen) Anaesthesize the area locally Spinal needle of 20 or 22 gauge is inserted into uterine cavity. 10-20 ml fluid is aspirated. AMNIOTIC FLUID COLLECTION 37

38. SAFETY PROCEDURES IN LABORATORY 38

39. Remain alert Be cautious while working 39

40. A fire extinguisher should always be handy. Keep sand bucket in the laboratory. Take measures to prevent electrical short circuiting. No smoking in the working zone of the laboratory. Breakable items should be kept in proper racks and never at the edge of the working table. Do not suck anything with the mouth, use rubber teets and bulbs for sucking. Do not place eatables on the working bench. Keep finger nails short PRECAUTIONARY MEASURES 40

41. At the end of the day clean all working benches with a disinfectant. See that nothing except the required electrical appliance is on. Dispose all infected material properly. The glasswares should be disinfected with a suitable disinfectant and be cleaned thoroughly with running water. Use rubber gloves and a nose mask while working. Wear a laboratory gown or uniform. 41

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43. Examples Storage Safe use Flammable chemicals Ether, xylene, ethanol, methanol, Romanowsky stain, acid alcohol etc. Cool storage In fireproof metal box At ground level Less quantity Never heat over flame Use water bath or electric hot plate Flame should be at least 10 feet away Corrosive chemicals Strong acids like conc. Sulphuric acid, nitric acid etc Strong alkalis like sodium hydroxide,potassium hydroxide Store them at low levels Never mouth pipette Pour at below eye level Always add corrosive substance to water slowly Types of chemicals : 43

44. Examples Storage Safe use Toxic, harmful & irritating chemicals Potassium cyanide, iodine, formaldehyde, chloroform, methanol, etc Store in locked cupboard Wear protective gloves Wash hands after using them Never mouth pipette Oxidising chemicals Chlorates, perchlorate, strong peroxide, chromic acid, etc Keep away from organic materials and reducing agents Keep away from flammable chemicals Handle with utmost care 44

45. Examples Storage Safe use Explosive chemicals Picric acid Store under water Do not allow to dry Never expose to flame Carcinogens Benzidine , nitrosamines, nitrosophenols , o- tolidine , o- dianisidine Level them as carcinogenic Handle with special precautions Wear protective gloves, mask, eye shields Do not allow contact with skin After use wash well with cold water 45

46. ACIDS ALKALIS TOXIC SUBSTANCES HEAT BROKEN GLASS ELECTRIC SHOCK CONTAMINATION BY INFECTED MATERIAL ACCIDENTS 46

47. Splashes on the skin : bathe the area with 5% aqueous sodium carbonate. Splashes in the eye : put 4 drops of 2% aqueous sodium bicarbonate into the eye. Swallowing of acid : Drink 5% soap solution immediately Gurgle with soap solution Give him 2 whites of egg mixed with 500 ml of milk or water 3 glasses of water Rinse the mouth and lips with 2% aqueous sodium bicarbonate ACID BURN WASH IMMEDIATELY WITH WATER 47

48. Splashes on the skin : bathe the area with 5% acetic acid/ vinegar Splashes in the eye : apply drops of saturated solution of boric acid Swallowing of alkalis : Make him drink 5% solution of acetic acid/lemon juice/diluted vinegar Gurgle with the same 3 glasses of water Rinse the mouth and lips with 5% acetic acid ALKALI BURNS WASH IMMEDIATELY WITH WATER 48

49. Send for a physician or qualified nurse Place the victim in the open air POISONING 49

50. Severe burns : If splashed with burning flammable solvent;roll him in a blanket or overall Lay the victim on t5he ground Cover him if he is cold Inform the physician Minor burns : Plunge the area with cold water or ice water. Apply mercurochrome or acriflavine ointment Apply dry gauze dressing If infected inform the physician. HEAT BURN 50

51. Wash the wound and remove the glass pieces. Apply mercurochrome or acriflavin ointment on the wound Cover with gauze and adhesive tape. If it bleeds profusely try to stop bleeding by pressing down on it and refer the patient to a physician. INJURIES CAUSED BY BROKEN GLASSES 51

52. Wash the wound. If it is not bleeding squeeze hard to make it bleed Bathe the area with antiseptic lotion. Wash thoroughly with soapy water. Bathe again with antiseptic lotion Refer the patient to the physician. If swallowed wash thoroughly with water and diluted antiseptic lotion CONTAMINATION BY INFECTED MATERIALS 52

53. It is rare Put off the main switch Send for a physician Begin mouth to mouth respiration immediately ELECTRIC SHOCK 53

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