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| Name | Class |
|---|---|
| University Medical Center Groningen | OTHER |
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Human milk oligosaccharides (HMOs), the third most abundant constituent of breastmilk, are known to have beneficial effects on infant immunity. Maternal genetic polymorphisms cause HMO variability. The FUT2 gene determines the secretor status, whereas the FUT3 gene is responsible for the expression of Lewis fucosyltransferase. Therefore, breastmilk can be classified to four groups according to the variation. To date, this variability has not been investigated in Indonesia. This study aims to evaluate the association between FUT2 gene polymorphism and 2'-Fucosyllactose (2'-FL) secretor phenotype. In addition, infant FUT2 gene polymorphism and short chain fatty acid (SCFA) profile from stool samples are also analysed.
Eligible mother-infant pairs are explained about this study. Those willing to participate in this study are asked for written informed consent. Mothers are interviewed about their baseline characteristics, family pedigree, nutritional intake, and routine drug consumption. Infants are checked for their birth history. Both are measured for weight and height.
Four specimens are collected from the subjects:
Mother
Breastmilk
Breastmilk is expressed at 8-11 AM to avoid variability due to circadian rhythm. One breast is emptied, 30 mL of breastmilk is stored in a sterile container, and the rest is returned for feeding. Breastmilk is divided to five 2-mL cryovials and stored in a -80°C freezer. The remaining is stored in a -20°C freezer.
Blood
Blood samples (3 mL) are collected for DNA extraction.
Infant
Buccal cells
Buccal swab kits are used to obtain samples for DNA extraction.
Stool
Stool specimens are collected at the age of four weeks in a sterile container and stored in a -80°C storage before short chain fatty acid profile analysis.
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| Measure | Description | Time Frame |
|---|---|---|
| Human milk oligosaccharides profile of Indonesian mothers | The concentration of 19 HMOs will be measured using high performance anion exhange chromatography with pulsed amperometric detection (HPAEC-PAD). | 1 day |
| Proportion of FUT2 secretor genotype of mothers based on single nucleotide polymorphism (SNP) rs601338 | Sequencing of coding sequence (exon 2) of FUT2 gene will be performed using a previously known outer primer from the study by Lefebvre, et al (2020). The results will be aligned with database from www.ncbi.nlm.nih.gov to find the proportion of rs601338 among mothers. | 1 day |
| Association between rs601338 FUT2 secretor genotype and 2'-FL secretor phenotype in mothers | The association between rs601338 FUT2 genotype (homozygous dominant, heterozygous, homozygous recessive) and 2'-FL concentration will be analysed using ANOVA test. | 1 day |
| Measure | Description | Time Frame |
|---|---|---|
| Novel FUT2 polymorphism in Indonesian mothers | The sequencing result of exon 2 in the FUT2 gene will be aligned with database from www.ncbi.nlm.nih.gov to find novel variants. Additional information regarding existing variants will be browsed using the same database. The allele frequency of new variants will be calculated to determine new SNP. | 1 day |
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Inclusion Criteria:
Mother
Infant
Exclusion Criteria:
Mother
Infant
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Eligible mothers and infants visiting the research site during recruitment.
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| Name | Affiliation | Role |
|---|---|---|
| Klara Yuliarti, MD | Dept of Pediatric, Faculty of Medicine, Universitas Indonesia/ Cipto Mangunkusumo General Hospital | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Faculty of Medicine, Universitas Indonesia | Jakarta Pusat | DKI Jakarta | 10440 | Indonesia |
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| ID | Term |
|---|---|
| D001942 | Breast Feeding |
| ID | Term |
|---|---|
| D005247 | Feeding Behavior |
| D001519 | Behavior |
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Breastmilk, whole blood, buccal cells, stool
| Proportion of FUT2 secretor genotype of infants based on single nucleotide polymorphism (SNP) rs601338 | Sequencing of coding sequence (exon 2) of FUT2 gene will be performed using a previously known outer primer from the study by Lefebvre, et al (2020). The results will be aligned with database from www.ncbi.nlm.nih.gov to find the proportion of rs601338 among infants. | 1 day |
| Novel FUT2 polymorphism in infant | The sequencing result of exon 2 FUT2 will be aligned with database from www.ncbi.nlm.nih.gov to find novel variants. Additional information regarding existing variants will be browsed using the same database. The allele frequency of new variants will be calculated to determine new SNP. | 1 day |
| Profile of infant stool short chain fatty acid based on mother-infant FUT2 genotype pairs | The concentration of total short chain fatty acid, acetate, propionate, and butyrate in infant's stool will be measured using gas chromatography-mass spectrometry (GC-MS). Difference in the concentration between four mother-infant genotype pairs will be analysed using ANOVA test. | 1 day |