VINIF.2023.DA061 – Establishment of Reference Intervals for Blood Biomarkers in Vietnamese Children and Adolescents

project manager image
Principle Investigator
Assoc.Prof. Tran Minh Dien
Host Organization
Vietnam National Children’s Hospital

Project goals

Establish a comprehensive database of reference ranges for blood test results in Vietnamese children and adolescents.

  1. Establish age- and sex-specific reference intervals for biochemical indicators in infants and children from 30 days old to 18 years old.
  2. Establish age- and sex-specific reference intervals for some endocrine markers and reproductive hormones in infants and children from 30 days old to 18 years old.
  3. Establish age- and sex-specific reference intervals for hematological and coagulation indices in infants and children from 30 days old to 18 years old.
  4. Establish continuous reference intervals for some biochemical indicators, some endocrine markers, reproductive hormones, hematological markers and common blood coagulation in infants and children from 30 days old to 18 years old.

Main tasks of the project

Select the study population

Children from birth to 18 years of age are eligible to participate in this research project, with written informed consent from their guardian. Sampling locations of 63 provinces and cities of Vietnam: with 7 geographical regions, 1 province will be selected for each region.
Through a community-based approach, events will be held at schools, community centers, kindergartens and special community clinics to create opportunities for children across Hanoi and some Provinces and cities across Vietnam to participate. Inclusion criteria for the study were determined by questionnaire, requesting information on chronic and acute medical history, prescription and non-prescription drug use, diet and ethnicity. Subjects were excluded from the study if they reported: 1) history of chronic or metabolic disease, 2) acute illness in the past month, and 3) prescription drug use in the past month.

Collect and analyze samples

Sample size: The expected sample size is at least 1680 healthy children, divided into 6 groups according to the child’s developmental stages, each group has at least 260 subjects, males and females in each group are equivalent. The six groups include: newborns (0-3 months), infants (4-11 months), toddlers (1-2 years old), preschoolers (3-5 years old), school age (6- 13 years old), teenagers and adolescents (14-18 years old).
Depending on the child’s age, 2 mL to 10 mL of whole blood will be collected in appropriate collection tubes depending on the test by professional pediatric phlebotomists trained in standard practice procedures. Blood samples for biochemical, immunological, and coagulation tests were centrifuged at 4000 rpm for 10 minutes within four hours after collection, divided into frozen vials of 0.5 mL or 1.5 mL and stored at -80°C. Blood samples for blood cell testing will be collected and performed within 24 hours of sampling.


35 analytes including electrolytes, metabolites, proteins, enzymes. 12 immunological indexes, endocrine indexes and reproductive hormones. 25 basic hematological parameters. 3 basic blood coagulation indexes.

Sample processing and sample analysis

Pre-test sample processing will be completed within four hours of collection. Samples will be transported refrigerated in batches to the laboratory and analyzed along with regular patient samples. Samples will be analyzed on testing systems according to the manufacturer’s instructions and include servicing, maintenance, calibration and quality control.

Data analysis and statistics

A direct normalization approach will be used to establish all reference intervals. Reference intervals were calculated based on CLSI guidelines (C28-A3). In parallel with the discrete reference intervals for each age group, continuous reference intervals will be established to accurately describe the fluctuations of biological markers for the developmental stages of children from birth to 18 years old.

project manager image
Principle Investigator
Assoc.Prof. Tran Minh Dien
Host Organization
Vietnam National Children’s Hospital


Expect Progress
Phase 1

– Present project proposal to the Central Pediatric Hospital’s Scientific Council.
– Submit project proposal for approval to the Hospital’s Ethics Committee.
– Develop comprehensive and detailed research plans for the entire project.
– Report to the Department of Science, Technology, and Training, Ministry of Health regarding the project and implementation plan.
– Review and evaluate the detailed project plan.
– Conduct team meetings with key project members.
– Procure equipment.
– Procure consumables.
– Coordinate with provinces/cities in the Northern, Central, and Southern regions to arrange sampling locations.
– Hold team meetings for laboratory research members.
– Draft standard operating procedures.
– Prepare interview questions, medical records of volunteers participating in the study, and Informed Consent Forms.
– Complete research proposal outlines.
– Review and approve standard procedures and developed forms.
– Train sampling staff.
– Train technicians, support staff, researchers, and postgraduate students.
– Collect samples at the Central Pediatric Hospital (children of healthcare workers volunteering for the study).
– Confirm methods, evaluate IQC, EQA results for all participating devices.
– Collect patient samples for method comparison.
– Conduct comparative method experiments.
– Evaluate pre-station at sampling sites (7 ecological zones, 1 province per zone).
– Collect samples in Northern provinces.
– Collect samples in Central and Southern provinces.
– Analyze blood formula samples.
– Analyze biochemical, immunological, and coagulation samples.
– Defend proposals of 3 doctoral students.
– Hold regular research team meetings every 3 months.
– Defend thesis proposals for graduate students.
– Write 2 papers and submit for publication in domestic journals.
– Organize training workshops for research team on statistical analysis and data processing.

Phase 2

– Analyze samples of biochemistry, immunology, and blood coagulation.
– Write and submit applications for publication of 2 papers in domestic journals.
– Analyze data and write a report, submit for publication in an international journal.
– Register to present results at an international conference.
– 3 research students report on project progress.
– Postgraduate students defend their theses.

Phase 3

– National workshop on pediatric reference ranges.
– Publishing papers domestically.
– Publishing papers internationally.
– Compiling printed and electronic books.
– National conference to announce project results.
– Presenting reports at international conferences.
– Graduate students defending their theses.