Tips for collection and handing of specimen

Blood

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Blood

1) Time of collection : As the test result may vary according to change of physiological conditions such as meal and exercies, it is general principle to collect blood on an empty stomach in the morning. For outpatients, collect at lest 2 hours after a meal. After a strenuous exercise rest enough before collectio. If the same test is repeated for a patient, it is desirable to collect under the same condition and at the same time.
2) Generally, when collecting blood at the same time for various tests (whole blood, plasma, serum, etc.), calculate the quantity of blood to be collected in advance. Please refer to the quantity of specimen by test item.
Whole blood Collect quantity required for the test
Serum (plasma) Collect about 3 times more than the required serum (plasma)
3) Do not apply Tourniquet for more than one minute, and collect the blood at a moderate speed to avoid bubbles. Concentration and hemolysis of blood may affect the test result.
4) Please check the specimen container by test item and collect it in the designated container. For continuous collection of blood in several tubes, please follow the "Blood collection sequence" as blow.

Collection sequence

5) In case of blood collection using a syringe, dispense the blood slowly so that the blood flows down the wall of the test container. (To avoid hemolysis)
6) Use an appropriate container for each anticoagulants it. As Silica particles are added inside wall of SST, shake the SSt like an anticoagulant tube.
7) Blood specimen should be collected and separated into serum or plasma (3000 rpm, for 10 minutes). The specimen should be refrigerated if it is difficult to collect on the same day.

Factors that cause errors in test result

1. Bacterial Test If the anticoagulant and the blood are not mixed sufficiently, the hematological test will result in inaccurate results due to partial clotting. Therefore, when collecting blood in a container with anticoagulant, mix it well (1-2 minutes) and request as soon as possible. 2. Hemolysis The reaction of hemoglobin with the intracellular substances due to hemolysis affects the result, causing increase of the value. therefore, attention should be paid when collecting and dispensing the blood. When collecting the blood with a syringe, do not pull the piston too hard and open the tube's cap and remove the needle to pour down slowly into the wall of tube. In addition, centrifuge the specimen after 20 minutes after the collection.

Test items which the results are affected by hemolyzed specimen

Increase Decrease
Total protein, K, Iron, Inorganic P, Amylase, AST, ALT, LDH, Cholesterol etc. Bilirubin, Lipase, APC etc.
3. Hyperlipidemia turbidity Blood collected right after a meal may be turbid of Such turbidity may affect the lipid test, resulting in correct result(Increase). Therefore, according to the specimen, the blood collection should be performed after 12 hours fasting.

Test items affecting the result of postprandial blood sampling

Increase Decrease
Amount of urine. glucose(blood, urin). Trglyceride. Uric acid. Amylase. Insulin. Cholesterol. Lipoprotein. Catecholamine. Aldosterone. VMA, Vitamin etc. Hb, Hct, WBC, Total protein, Albumin, Iron etc.

Urine test

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Urine Test

1) Random urine : Take the medication urine of the first urine in the morning and request as soon as possible. Please refrigerate the specimen until transportation.
2) 24 hours urine : Urinate and discardat 08:00 in the morning. Collect 24 hours urine untill 08:00 next morning in a collection bag (Include urine at stool). Most the specimen can be stored in refrigerated condition. However, if it is mandatory to add preservatives (refer to next page), first add it into the collection bag to collect the specimen. Mix well about 10ml of the 24 hours urine in a container for request (Fill in the volume of urine)

Body fluid test

Pleural Ascitic, and Joint Fluid. which are easily coagulated after collection (excluding CSF), may cause a large error in cell count or chemistry test items when coagulated.

Specimen container Test item
EDTA tube Body fluid analysis, Some of Chemistry items
Plain tube Chemistry items(If not clotted). Cytological test

Available urine preservative by test item

Test name 24 hours urine preservative
No preservative Toluene 6N HCL 50% Acetic acid Boric acid Na2 CO3 HNO3
17-Kestosteroids 체크표시 체크표시
17-OHCS 체크표시 체크표시
5-HIAA* 체크표시 체크표시 체크표시 체크표시 체크표시 체크표시
Alodosterone 체크표시
Aluminum(Al) 체크표시 체크표시 체크표시 체크표시 체크표시
Arsenic(As) 체크표시 체크표시 체크표시 체크표시 체크표시 체크표시
RUN 체크표시
Cadmium(Cd) 체크표시 체크표시 체크표시
calcium(Ca) 체크표시 체크표시 체크표시
Catecholamine* 체크표시
Chloride(Cl) 체크표시
Citrate 체크표시
Cobalt(Co) 체크표시 체크표시
Copper(Cu) 체크표시 체크표시 체크표시 체크표시 체크표시 체크표시
Cortisol 체크표시
Creatine 체크표시 체크표시
Creatinine 체크표시 체크표시 체크표시
Cyclic AMP(cAMP) 체크표시
HVA(Homovanilic acid)* 체크표시 체크표시
Hydroxyproline, free 체크표시 체크표시
Hydroxyproline, total 체크표시 체크표시 체크표시 체크표시 체크표시 체크표시
Lead(Pb) 체크표시 체크표시 체크표시 체크표시 체크표시 체크표시
Magnesium(Mg) 체크표시 체크표시
Mercury(Hg) 체크표시
Metanephrine* 체크표시
Microablumin 체크표시
Osmolality 체크표시
Oxalate 체크표시
Porphobilinogen 체크표시
Potassium(K) 체크표시
Protein E.P 체크표시
Protein, total 체크표시 체크표시
Uric acid 체크표시 체크표시
VMA(Vanillymandelic acid)* 체크표시
Zinc(Zn) 체크표시 체크표시 체크표시 체크표시 체크표시 체크표시
δ-ALA 체크표시 체크표시
(*:Preservative essential, 체크표시 : Recommended, 체크표시 : Possible)

Body fluid test

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Body fluid test

Body fluid : Pleural, Ascitic, and Joint Fluid, which are easily coagulated after collection (excluding CSF), may cause a large error in cell count or chemistry test items when coagulated.
  • Use of Edta bottle : Some of Chemistry items
  • Use of Plain tube : Chemistry item (If not clotted). Cytological test
  • Use of Heparin tube : when detecting crystal in join fluid

Microbiological test

  • 1.Refer to the quantity of specimen listed in this test guide.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Microbiological test

General precautions for microbiological test

1. General Bacteriology test

  • · Fill in the clinical findings, suspected organisms, the type of specimen, the time of collection, and whether an antibiotic was administered in a request form. Further more make sure not to miss the information as the test operation differs according to the type of specimen and location of sampling.
  • · Microbial specimens should be collected in a designated sterile container, and the operation should be performed aseptically.
  • · Keep BHI in the refrigerator and Thioglycollate broth at room temperature. Thioglycollate broth should be replaced if the purple layer at the top of Tioglycollate broth occupies more than 1/3.
  • · Use the Transport Medium for swab samples (ex. pus, discharge, etc.). When requesting a Gram stain at the same time, take it on two swabs and put it in a transport medium.
  • · All bacteriological specimens should be sent to our laboratory immediately after collection but keep them in a refrigerator in an unavoidable case.
  • · Specimen must be kept at room temperature for Meningococcus, Gonococcus culture.
  • · In case of urine culture test, the colony is counted. Therefore, please note that if specimen collection is not appropriate there is no need to do the test. (Clean the urine tool well, then take the medium urine, and keep it refrigerated)
  • · The date of report is as follows.
  • Blood culture: 3 ~ 7 days (Final report takes 7 days)
  • Culture except blood: 2 ~ 3 days later
  • Culture of Mycobacterium tuberculosis: Interim report after 4 weeks (Final report 8 weeks)
  • Fungal culture: Interim report every week (Final report 3 Weeks)

2. Mycobacterium tuberculosis test

  • Sputum: Take the first sputum once in the morning. It is advisable to repeat the procedure for 3 ~ 5 days before the treatment begins. (Completely seal the container to prevent the specimen from flowing out of the container)
  • Urine: Take the first urine in the morning. 24 hours urine is not suitable.
  • Gastric juice:Collect after fasting for over 8 hours.
  • Body fluid: Collect asptically like general bacterial test

Precautions for each specimen

1. Urine

  • · Midstream urine, urine from catheter and suprapubic aspiration urine are used for bacterial culture.
  • · The best specimen is the midstream urine, of the first urine in the morning. Wash the surroundings of urethra with soap water and then discard the first few ml and take 10ml of medium urine in an aseptic container.
  • · Catheter urine may cause a nosocomial infection. Therefore, it is performed only for the patient who cannot urinate or who requires aseptic operation.
  • · Suprapubic aspiration is performed for the infant who is difficult to collect the urine or in the case of suspected anaerobic bacteria.
  • · Foley Catheter tip is not cultured as the of urethra is contaminated with normal flora.
  • · 24 hours urine is not cultured.
  • · Refrigerate the specimen if it is delayed over 2 hours.

2. Wound infection and abscess

  • · Purulent specimen collected with a syringe is recommended but a swab specimen taken from deep part of the lesion is acceptable.
  • · Generally, two swab specimens are required for bacterial culture.
  • · Refrigerate when delayed, but keep at room temperature when bacteria are anaerobically cultured

3. Sputum, Tracheal aspirate and Throat

  • · Rinse the mouth with water and cough deeply before breakfast early in the morning, and take 5~10ml of sputum.
  • · In case of having dry cough, inhale spray with 15% saline and 10% glycerin to induce sputum.
  • · If the specimen contains saliva as determine the quality follows.

<Murray and Washington's grading system for assessing the quality of sputum samples>

Grade Epithelial cell WBC
1 >25 <10
2 >25 10-25
3 >25 >25
4 10-25 >25
5 <10 >25
6 <10 <10
★(Clinical Microbioloby Procedure Handbook. Washington DC: ASM, 2004: 3.2.1-3.2.1.19)
Grade 1-3,6 : Unacceptable
grade 4-6 : Acceptable
  • · Put a cotton swab between tonsils and mucous membranes so that they do not reach the oral mucosa or the mucous membranes while pressing tongue with tongue depressor and ask the patient to pen the mouth with "ah" sound and gently rub the posterior pharynx or inflammatory tonsils to collect the specimen
  • · If delivery is delayed, keep it in refrigerator (The specimen to detect Bordetella pertussis at room temperature)

4. Stool

  • · The stool specimen is good for Bacteriological test related to infectious diarrhea, but can be replaced with rectal swab specimen.
  • · Collect the specimen in a clean container with a good stopper to avoid contamination by urine or toilet paper.
  • · Collect the specimen which contains pus, blood and mucus.

5. Blood

  • · Wait for one to two minutes to dry after thoroughly disinfecting the vascular skin with iodophor, iodine tincture, and chlorhexidine to collect the blood.
  • · It is best to collect before having the chills but since the time is unpredictable.. Therefore, take immediately after fever and collect specimen 2~3 times from other areas at the same itme.
  • · Do not collect blood only once from adult.
  • ·Blood:The appropriate ratio of the medium to blood is from 1:5 to 1:10. Usually, 10 ml of blood is collected from the vein once, and then the same amount is divided into the medium for aerobic and anaerobic culture to inoculate.
  • · For children, take 1 to 5 ml of blood (No more than 1% of total blood volume) to inoculate the aerobic culture bottle.
  • · the inoculated blood culture bottles should be kept at room temperature until they arrive at the laboratory.

6. Genitourinary tracts

  • · Get into the cervix in one to two seconds and rub the wall for 10~30 seconds to collect and put it in a transportation medium.
  • · Don't store the specimen to identify Neisseria gonorrhea in a refrigerator but at room temperature.
  • · To diagnose C. trachomatis or gonorrhoeae in a male, put a cotton swab 2~4 cm into the urethra at least 2 hours after urination and rotate for about 6 seconds to collect the urethral test specimen.

7. Body fluid

  • · For Cerebrospinal fluid, use the specimen from the second aseptic tube after skin disinfection as blood culture for gram stain and culture. (First aseptic tube: biochemical and serological test, third aseptic tube: for cell count)
  • · Collect at least 0.5ml of Cerebrospinal fluid and make sure to keep at room temperature.
  • · Take body fluids such as pleural fluid, ascites, pericardial fluid, and joint fluid with a syringe and sterilize the skin in the same way as blood culture to collect specimen.
    Sterilize the skin in the same manner as blood culture and collect specimens.

8. Catheter tip

  • · Intra-arterial or intravenous catheter
  • · Intravenous catheter tip should be sterilized with 70% alcohol, aseptically removed, cut into 2 ~ 3 inches at the end and transported in aseptic screw cap tube. If delivery is delayed, it should be kept in a refrigerator.
  • · Indwelling urethral catheter tip, catheter tip in a saline or transportation medium, catheter tip smeared on a slide are not suitable for culture.

9. Anaerobic

  • · After aspiring with a syringe, bend the needle and cover with a rubber stopper to prevent exposure to the air.
  • · Pharynx, nasopharynx, oral mucosa, sputum, bronchial washing solution, contents of stomach and intestine, stool, rectal swab, urine collected by catheter, swab specimen from vagina and cervix, cerebrospinal fluid, and specimens in the surface of bed sore and mucous membrane are not suitable for anaerobic bacteria culture.

10. Fungal test

  • · If the KOH mount specimen is skin, hair or keratin, the potassium hydroxide dissolves the keratin in these specimens to facilitate the observation of fungi, because the cell fragments cover the fungal particles.
  • · After cleaning the keratinous area with 70% alcohol and scraping its outer layer with a sterilized blade and scratching the specimen from the infected area inside the keratin, wrap it with paper and put it in a sterilized container to request.
  • · First, wipe the skin with 70% alcohol and remove the bacterial flora. Then, if there is a ringworm, scrap out the outer part of red circle. If there is no red circle, scrap out the most infected area.
  • · In order to obtain the most suitable specimen, aseptically collect the most damaged lesion before the antimicrobial treatment.
  • · Take sufficient quantities and repect sampling as much as possible

11. Mycobacterium tuberculosis test

  • · Sputum
  • - Collect the first morning sputum once in a sterile container.
  • - It is recommended to repeat the test for 2-3 consecutive days
  • - Swab specimen is not appropriate for the cultivation of acid-fast bacterium. Due to the hydrophobicity of the acid-fast bacterium, migration from the swab specimen to the medium is not easy, resulting in limited volume of specimen.
  • - Pooled sputum, which is a collection of sputum, is not suitable due to increased contamination.
  • · Urine
  • - Collect the midstream urine of the first morning urine in an aseptic container for 2~3 consecutive days
  • - For 24 hours urine, a small amount of urine is unsuitable.
  • - Collection with a catheter is only limited when the midstream urine cannot be obtained.
  • · Body fluid
  • - Use Sodium polyanethol sulfonate (SPS) anticoagulant to collect the blood containing fluid.
  • - At least 5ml of cerebrospinal fluid is needed as it contains only a few acid-fast bacteria.
  • · Tissue, pus, etc.
  • - Specimen exposed to formalin is unsuitable for smearing and culture.
  • - Tissues should not be placed in saline or wrapped with gauze, but the microscopic biopsy specimens can be transported in small quantities of sterile saline.

12. Virus test

  • · Use a designated container for virus test as the microbial contamination may affect virus isolation.
  • · Keep specimen for virus test refrigerated immediately after collection. (However, keep blood specimen at room temperature)
  • · Aseptically collect and put in a sterile container to request
  • · Record the clinically suspected viruses and site of sampling together.

Test for congenital metabolic
abnormalities test

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Test for congenital metabolic abnormalities

  • Time for blood sampling : The recommended blood sampling period is between 48 hours and 1 week after birth.
    In the case of premature infant and low birth weight infant, blood sampling is performed 5 to 7 days after birth.
    Later, when the feeding condition improves, collect the blood specimen again.
    If the feeding condition does not improve, it is necessary to collect 5 to 7 days and 1 month after birth.
  • Blood sampling paper : Do not touch or contaminate the sampling area of the filter paper. If something is written on this area with ink or pen or it is stained with cosmetics or amino-acid, screening test may be inaccurate. Therefore, it must be kept in good condition. Handling the paper after the sampling.
  • Avoid direct sunlight or hot places after sampling and dry naturally in a cool and dark place
  • Allow to dry thoroughly for at least 3 to 4 hours

Cytopathology test

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Cytopathology test

Precautions on gynecological test

1. Do not use lubricant or talcum powder on instruments or gloves when sampling the specimens. Use warm saline to add moisture.
2. Do not perform biopsy or induce other trauma before sampling. (Dilution or masking of blood may interfere with diagnosis)
3. If swap is used for sampling, take the specimens from the vaginal fornix and cervix and smear them thoroughly on a slide.
4. When sampling is made with Pap brush or Cytobrush, avoid excessive bleeding (it may interfere with diagnosis)
5. Refrain from intravaginal medication and do not use contraceptive pill at least 24 hours prior to sampling
6. To prevent detached cells from deteriorating, be sure to fix them immediately (within 2 seconds) after thinly smearing them on the cleaned surface of a slide. (If it is dried in the air after smearing, it causes denaturation of cells and diagnosis is impossible)
7. If a malignant tumor is suspected clinically, but the result is negative, repeat or perform biopsy because sampling and smear fixing may have been inappropriate.
8. Cytological test may lead to difficulty in diagnosis due to lack of prior knowledge and careless handling when sampling, storing and fixing of specimen. For precise diagnosis, please make sure to carefully read the precautions such as sample collection, fixation and storage condition before sending the specimen for test.

Smear sampling of cell specimen and how to request (process)

  • 1. Gynecological test
    • · Smear immediately in 2 seconds on a slide after sampling from uterine cervix. Fix tightly with Cytofixer or fix it in 95% Ethyl alcohol for more than one hour.
  • 2. Sputum
    • · Request with patient's sputum not saliva.
  • 3. Accumulated sputum (for 3~5 days)
    • · Take the first sputum of the morning once a day and by sampling day
  • 4. Take body fluid as much as possible (More than 10ml) (Pleural & Ascitic fluid, Urine, Bronchial washing etc.)
  • 5. FNA (Find Needle Aspiration)
    • · Immediately smear the aspirated specimen on a slide (multiple sheets, if possible) and fix it in 95% Ethyl Alcohol for more than one hour and send us the remaining specimens.

Histopathology test

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Histopathology test

  • · Normally, the specimen (biopsy or extracted tissus) is fixed with 10% formalin.
  • · The fixative is put 10-15 times of tissue and placed at room temperature for 12-14 hours.
  • · Place the tissue sampled from different part of the tissue to be distinguished in a separator container.
  • · Stomach biopsy tissue: Attach the biopsy tissue to the filter paper by number and fix it in a bottle containing the fixative.
  • · Avoid unnecessary actions on the extracted tissues and put sufficient amount of fixative so that they do not dry out after extraction.
  • · Please use a specimen container with a wide opening and list the patient's personal details and the client's name on the back of the container.
  • · Please include the patient's name, gender, age, and requested hospital, date of collection, sampling site, clinical diagnosis, medical history and surgical findings of the clinician.

Prerequisite to histopathology request

1. Please fill in the hospital name, patient's name, age, gender and chart number on the container.
2. Please make sure to fill in the hospitals name, patient's name, age, gender, date of collection, clinical diagnosis, medical history and previous biopsy and especially clinical findings, because they are very important for accurate diagnosis.

Cytogenetic test

  • 1.Refer to the quantity of specimen listed in this test guide. If it is stored the urine (24 hours urine),
    please send only the required quantity in the container and make sure to write the total quantity of urine.
  • 2.Please collect specimen in the designated container. If a special container is needed, ask for it in advance before the collect the specimen.
  • 3.If the specimen is smeared on a side, please write the name, gender and age with a bandage and a pencil.
  • 4.To request a consecutive test, be sure to write the time of consecutive test and the name of medication taken.

Cytogenetic test

  • 1. Precautions on Requesting Chromosome Test
    • · Amniotic fluid : Discard the first two to three drops to prevent the maternal cell contamination
    • · If specimen contains excessive blood or the color of amniotic fluid is brownish, cultivation may fail. Therefore, monthers should be fully informed in advance.
    • · conical tube or heparin bottle is commonly used for the specimen. Wrap and seal the specimen with a foil when sending the syringe together.
    • · Specify the gestational age and the purpose of the test on the request form.
  • 2. Chorion
    • · Collect the specimen in an exclusive container provided by the laboratory.
    • · If only maternal cells without Villi are collected, the Cytogenetic Team will observe them under a microscope and notify the client through the phone for rechecking.
  • 3. Aborted tissue
    • · Collect the specimen in an exclusive container containing the phosphate buffer supplied by the laboratory.
      (If a specimen is treated with formalin, chromosome test cannot be performed.)
    • · the specimen collected from the chorion and skin tissus (size: W x L 1cm) has a high success rate.
    • · Because aborted tissues are easily contaminated with high rate of culture failure, it is necessary to sufficiently explain to the caregiver in advance.
  • 4. Blood and bone marrow
    • · Take 2 ~ 3ml of specimen in a sodium heparin bottle and pay attention to avoid contamination and coagulation.
    • · The clinical information of the patient is necessary because the culture method may vary.
  • 5. FISH test
    • · For the test of chromosomal abnormity, please specify the chromosome number. In case of prenatal test, it is possible only when requested with the cultivation method
    • · For diagnosis of microdeletion disease, detailed clinical information of the patient as well as its disease name is required.
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