Blood and Serum Samples

  1. Labelling Blood Collection Tubes
    1. Affix the patient label in a straight line on the tube; do not wrap it around in a spiral.
    2. Ensure the tube color code and fill line remain visible after the label is applied.
    3. If the sticker is longer than the tube, trim the excess—keeping the HN and patient name visible—or fold the extra portion back on itself.
  2. Blood Collection Guidelines
    1. Verify that the patient name on the request form matches the label on the blood collection tube.
    2. Ask the patient’s full name before every blood draw.
    3. Avoid applying the tourniquet for more than 1 minute to prevent alteration of test results. If vacuum collection is not used, do not remove the cap—instead, pierce through the cap with the needle and allow the vacuum to draw the blood without pushing the syringe plunger.
  3. Multiple Tube Collection: Order and Mixing Instructions
    1. Clotted blood tube: Invert 5 times
    2. EDTA tube: Invert 8 times
    3. Heparin tube: Invert 8 times
    4. Citrate tube: Invert 8 times
    5. Plasma or serum must be separated by centrifugation immediately after blood collection.
  4. Eligible Submitters – Patients from the Hospital for Tropical Diseases, other hospitals, research institutions, and private companies may submit specimens in appropriate collection tubes according to the specimen type (see table below).
Specimen Types, Volumes, and Storage Conditions
No. Test Specimen Type Volume Storage Conditions
1 Detection and serotyping of Dengue virus using Real-time RT-PCR EDTA blood, Clotted blood, EDTA plasma, Serum Adults and children >12 years: 2 mL Children <12 years: minimum 1 mL 2–8 °C for up to 6 hours If delayed, separate serum or plasma and store at −20 °C to −80 °C
Collect at least 0.5–1 mL of plasma or serum in a microcentrifuge tube. Ensure the cap is tightly closed and sealed with parafilm.
2 Detection of anti-Dengue virus IgM and IgG antibodies using ELISA test EDTA blood, Clotted blood, Citrate blood, Heparinized blood, Plasma, Serum ผAdults and children >12 years: 2 mL Children <12 years: minimum 1 mL 2–8 °C for up to 24 hours If delayed, separate serum or plasma and store at −20 °C to −80 °C
Collect at least 0.5–1 mL of plasma or serum in a microcentrifuge tube. Ensure the cap is tightly closed and sealed with parafilm.
3 Detection of Chikungunya virus using Real-time RT-PCR EDTA blood, otted blood, EDTA plasma, Serum EDTA plasma: 2 ml, EDTA blood 5 ml, Clotted blood 5 ml, Serum 2 ml 2–8 °C for up to 24 hours If delayed, separate serum or plasma and store at −20 °C to −80 °C
4 Detection of Zika virus using Real-time RT-PCR Urine, EDTA blood, Clotted blood, EDTA plasma, Serum Urine 2 ml, EDTA plasma 2 ml , Serum 2 ml, EDTA blood 5 ml, Clotted blood 5 ml 2–8 °C for up to 24 hours If delayed, separate serum/plasma/urine and store at −20 °C to −80 °C
5 Diagnosis of Parasites Using the Stool Concentration Technique Stool sample Collect about a thumb-sized amount of stool, sampling from different areas, especially those with color variations or abnormal characteristics. Stool samples collected at home should be submitted to the laboratory within 4 hours. If required, they can be stored in a regular refrigerator for up to 1 day.
6 Diagnosis of Parasites Using the Direct Wet Smear Technique Stool sample Collect about a thumb-sized amount of stool, sampling from different areas, especially those with color variations or abnormal characteristics. Stool samples collected at home should be submitted to the laboratory within 4 hours. If required, they can be stored in a regular refrigerator for up to 1 day.
7 Detection of Pinworm Using the Scotch Tape Technique Pinworm eggs in the perianal region Collect a perianal specimen by applying clear adhesive tape immediately after the patient wakes up, before bathing. Transfer the tape onto a microscope slide for examination Slides can be stored at room temperature for no longer than 24 hours.
8 Malaria Diagnosis Using Thick and Thin Blood Films EDTA Blood – Collect 10 µL of blood by finger-prick from the patient. – Collect 2–3 mL of EDTA blood. Specimens can be stored at room temperature for no longer than 24 hours.
9 Detection of Occult Blood in Stool Using Immunochromatographic Test Stool sample – Collect approximately a thumb-sized portion of stool, making sure to sample from throughout the stool mass. Select areas with different colors or any abnormal portions, such as those containing mucus or blood. – Use freshly passed stool, collected directly into the toilet or a clean container. – Avoid consuming the following foods and supplements 48–72 hours prior to stool collection: beetroot, carrots, bananas, grapes, cantaloupe, cucumber, broccoli, cauliflower, horseradish, turnip, various mushrooms, red meat, fish, chicken, iron supplements, pig’s blood, chicken blood, and vitamin C or vitamin C-containing beverages. Stool samples collected at home should be submitted to the laboratory within 4 hours. If required, they can be stored in a regular refrigerator for up to 1 day.
10 Detection of Zika, Chikungunya, and Dengue viruses using Multiplex Real-time RT-PCR EDTA blood, Clotted blood ผAdults and children >12 years: 2 mL Children <12 years: minimum 1 mL 2–8 °C for up to 24 hours If delayed, separate serum or plasma and store at −20 °C to −80 °C
Collect at least 0.5–1 mL of plasma or serum in a microcentrifuge tube. Ensure the cap is tightly closed and sealed with parafilm.

Specimen Collection and Handling for COVID-19 Diagnosis (SARS-CoV-2 by Real-time RT-PCR)

To ensure accuracy and reliability in laboratory testing, specimen collectors are advised to follow these guidelines:

  1. Preparation Prior to Sample Collection:
    • Prepare all necessary materials, including labeled collection tubes.
    • Ensure patient details (name, specimen type, and date) are clearly written on waterproof labels.
    • Wear appropriate PPE.
  2. Timing of Collection:
    • Collect specimens as soon as possible, preferably within 2–14 days after symptom onset, or at the latest, 5 days after exposure to the disease/patient.
  3. Lower Respiratory Tract Specimens (for severe cases with pneumonia or lung inflammation):
    • Collect specimens such as bronchoalveolar lavage, tracheal aspirate (2–3 mL), or sputum (0.5–1 mL) in sterile containers.
    • Do not use UTM/VTM unless the patient is intubated. In such cases, cut the end of the ET-tube and place it in the transport medium.
    • Also collect upper respiratory specimens to increase detection sensitivity.
  4. Upper Respiratory Tract Specimens (for mild cases):
    • Acceptable samples include nasopharyngeal aspirate, wash, swab, and throat swab.
  5. Swab Collection Guidelines:
    • Collect both nasopharyngeal and throat swabs and place in the same tube with 1–3 mL UTM/VTM.
    • Use Dacron or Rayon swabs with plastic or wire shafts (no calcium alginate).
    • Insert swab into the tube, break off the shaft, and seal the tube tightly.
  6. Sample Handling and Decontamination:
    • Use leak-proof tubes. Seal with tape after closing.
    • Change outer gloves and disinfect tube surfaces with 0.1% sodium hypochlorite.
  7. Storage and Transport:
    • Store samples in an icebox or refrigerator at 2–8 °C.
    • Deliver to the lab within 24 hours.
    • If delayed, freeze at -70 °C or lower

 

Specimen Packaging for Laboratory Submission

  1. Wrap the specimen tube:
    • Use enough absorbent material to soak up all liquid in case the inner tube breaks or leaks.
  2. Double-bag the specimen:
    • Place the tube in the first zip-lock bag and wipe the outside with 70% alcohol.
    • Put on a new pair of gloves and place the first bag into a second zip-lock bag. Wipe the outside again with 70% alcohol.
  3. Place in a sturdy plastic container (secondary packaging):
    • Must prevent leaks, have a tight-fitting lid, resist impact, and withstand disinfectant spraying.
  4. Place in a foam box (tertiary packaging):
    • Use a strong, shock-resistant foam box.
    • Put ice packs between the foam box and plastic container to maintain 2–8 °C during transport.
    • Seal the box with tape and spray the exterior with disinfectant.
  5. Label the box clearly:
    • Include: recipient’s name, address, phone number; transport supervisor’s contact; sender’s name and phone number.
    • Attach the Biohazard symbol and arrows showing the upright position.
  6. Dispose of PPE safely:
    • Remove gloves, gown, or other PPE and discard in a biohazard waste container for proper disposal or decontamination.

Mpox Virus Detection by Real-time PCR

To ensure the accuracy and reliability of laboratory diagnostics, specimen collection for Monkeypox virus detection via Real-time PCR should follow these guidelines:

  1. Preparation and PPE
    • Before collecting specimens, prepare all necessary materials, including specimen tubes labeled with patient details (name, specimen type, date) and cover the label with waterproof material.
    • Wear full PPE: liquid-impermeable gown, hair cover, N95 mask or PAPR (powered air-purifying respirator), face shield, double gloves, and shoe covers, or follow the PPE requirements of the healthcare facility.
    • After specimen collection, replace all PPE before entering other areas.
  2. Collection of Vesicle or Pustule Fluid
    • Clean the lesion with 70% alcohol.
    • Using a sterile syringe and needle, collect 0.5–1 mL of fluid from vesicles or pustules.
    • Transfer to a sterile tube, close the lid tightly, and store at 2–8 °C until transport.
    • If multiple lesions are available, collect fluid from several lesions into the same tube to ensure sufficient volume for virus isolation.
  3. Swab from Lesions
    • Clean the lesion with 70% alcohol.
    • Use sterile scissors to remove the skin covering the lesion, then scrape the lesion with a sterile lancet until the surface is moist but not bleeding.
    • Apply a sterile swab to the lesion and immediately place it into 1–2 mL viral transport media (VTM) or a sterile tube.
    • Break off the swab handle to close the tube tightly.
    • Store on ice (2–8 °C) immediately.
    • For increased viral yield, swab 2 sites or use 2 swabs and place both into the same VTM tube.
  4. Throat Swab
    • Collect from patients with lesions in the buccal mucosa or from close contacts of confirmed patients who are asymptomatic.
    • Swab the posterior pharynx and place the swab into VTM.
    • Break off the swab handle to close the tube tightly.
  5. Swab Material
    • Do not use calcium alginate swabs or wooden shaft swabs, as they may inhibit certain viruses or PCR reactions.
    • Use Dacron or Rayon swabs with wire or plastic shafts.
  6. Specimen Storage
    • Lesion fluid and swab samples must be refrigerated at 2–8 °C or frozen at -20 °C or lower within 1 hour of collection.
    • If testing cannot be performed within 1 week, store specimens at -70 °C or lower.

 

Specimen Packaging for Laboratory Submission

  1. Wrap the specimen tube:
    • Use enough absorbent material to soak up all liquid in case the inner tube breaks or leaks.
  2. Double-bag the specimen:
    • Place the tube in the first zip-lock bag and wipe the outside with 70% alcohol.
    • Put on a new pair of gloves and place the first bag into a second zip-lock bag. Wipe the outside again with 70% alcohol.
  3. Place in a sturdy plastic container (secondary packaging):
    • Must prevent leaks, have a tight-fitting lid, resist impact, and withstand disinfectant spraying.
  4. Place in a foam box (tertiary packaging):
    • Use a strong, shock-resistant foam box.
    • Put ice packs between the foam box and plastic container to maintain 2–8 °C during transport.
    • Seal the box with tape and spray the exterior with disinfectant.
  5. Label the box clearly:
    • Include: recipient’s name, address, phone number; transport supervisor’s contact; sender’s name and phone number.
    • Attach the Biohazard symbol and arrows showing the upright position.
  6. Dispose of PPE safely:
    • Remove gloves, gown, or other PPE and discard in a biohazard waste container for proper disposal or decontamination.