Best Practices for Human Milk Collection for COVID-19 Research

McGuire, Michelle K.; Seppo, Antti; Goga, Ameena; Buonsenso, Danilo; Collado, María Carmen; Donovan, Sharon M.; Müller, Janis A.; Ofman, Gastón; Monroy-Valle, Michele; O'Connor, Deborah L; Pace, Ryan M.; Perre, Philippe Van de

doi:10.1089/bfm.2020.0296

Cited by 25 publications

(21 citation statements)

References 69 publications

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“…Furthermore, heterogeneity, lack of standardisation of sample collection and processing within and between studies is a known challenge in breast milk research (64)(65)(66). The studies included in this review were no exception and, like others before us, we identified a need for greater standardisation and clearer reporting of sample collection and processing methods which we discuss below (64,66). Looking beyond the methodological limitations and challenges, we first discuss the current state of knowledge regarding the general miRNA profile in human milk, their stability and function and factors which affect them.…”

Section: Discussionmentioning

confidence: 99%

“…This is a common feature for young and fast-growing research fields as the coherence in methods and other study features are continually developing. Furthermore, heterogeneity, lack of standardisation of sample collection and processing within and between studies is a known challenge in breast milk research ( 64 – 66 ). The studies included in this review were no exception and, like others before us, we identified a need for greater standardisation and clearer reporting of sample collection and processing methods which we discuss below ( 64 , 66 ).…”

Section: Discussionmentioning

confidence: 99%

“…Heterogeneity, lack of standardisation of sample collection and processing within and between studies is a common challenge in breast milk research (64)(65)(66). The studies included in this review were no exception and, like others before us, we identified a need for greater standardisation and clearer reporting of sample collection and processing methods (64,66). There is currently insufficient evidence to determine if it is particularly important to standardise certain aspects of the collection and processing procedures when analysing ncRNAs, such as the time of day for collection, volume of milk, or the speed and duration of centrifugation.…”

Section: Methodological Considerations Within Studiesmentioning

confidence: 99%

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Non-Coding RNAs in Human Breast Milk: A Systematic Review

et al. 2021

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Breast milk is the primary source of nutrition and hydration for the newborn infant but also plays an important role in the child’s first immune defense. Additionally, several breast milk factors have been implicated in immune-related health outcomes later in life, including immunoglobulins, cytokines, chemokines, growth factors and, more recently, non-coding RNA (ncRNA) species. In this systematic review, we provide a comprehensive summary of the current literature on endogenous ncRNAs found in human breast milk. Thirty (30) relevant studies were identified and, whilst the majority studies focused on microRNAs (miRNAs), there is evidence that breast milk contains high quantities of RNA which also include long-coding RNAs, circular RNAs, as well as other short RNAs and fragmented tRNA and rRNAs. Among studies investigating miRNAs, miR-148a-3p, miR-30a/d-5p, miR-22-3p, miR-146b-5p, miR-200a/c-3p, and the 5p end of the let-7 miRNAs were commonly reported among the top 10 miRNAs in the cell, lipid, and skim milk fractions of breast milk. Methodological difference and small sample sizes limit the possibility of conclusively identifying which maternal and infant characteristics affect the miRNA profile. The highly expressed miRNAs were generally reported to be similar across lactational stage, milk fraction, maternal and infant characteristics, or infant growth and health. All the same, individual studies identify potential differences in miRNA expression levels which should be confirmed by future studies. Stability, uptake, and physiological functions of miRNAs were also considered in several studies. Breast milk miRNAs are relatively resistant to a range of harsh conditions and uptake experiments suggest that extracellular vesicles containing miRNAs and circular RNAs can be taken up by intestinal epithelial cells. Although the evidence regarding the functional effect of breast milk miRNAs is limited, the predicted functions range from metabolic and biosynthetic processes to signaling pathways, cellular adhesion, communication, growth, and differentiation. Finally, this systematic review highlights some of the methodological challenges and knowledge gaps which can help direct future research in this field. In particular, it is important to further investigate the bioavailability of miRNAs in different milk fractions, and to characterize other ncRNAs which are largely unstudied.Systematic Review RegistrationPROSPERO https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=138989, identifier CRD42020138989.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodological Considerations Within Studiesmentioning

confidence: 99%

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Non-Coding RNAs in Human Breast Milk: A Systematic Review

et al. 2021

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“…Breast milk was collected following a standardised protocol described elsewhere. 14 Details on collection, sampling and storage are described in online supplemental text S1 . Whole milk was used for SARS-CoV-2 RNA detection and whey milk was used for antibody determination.…”

Section: Methodsmentioning

confidence: 99%

SARS-CoV-2 RNA and antibody detection in breast milk from a prospective multicentre study in Spain

Bäuerl

Randazzo

Sánchez

et al. 2021

Arch Dis Child Fetal Neonatal Ed

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ObjectivesTo develop and validate a specific protocol for SARS-CoV-2 detection in breast milk matrix and to determine the impact of maternal SARS-CoV-2 infection on the presence, concentration and persistence of specific SARS-CoV-2 antibodies.Design and patientsThis is a prospective, multicentre longitudinal study (April–December 2020) in 60 mothers with SARS-CoV-2 infection and/or who have recovered from COVID-19. A control group of 13 women before the pandemic were also included.SettingSeven health centres from different provinces in Spain.Main outcome measuresPresence of SARS-CoV-2 RNA in breast milk, targeting the N1 region of the nucleocapsid gene and the envelope (E) gene; presence and levels of SARS-CoV-2-specific immunoglobulins (Igs)—IgA, IgG and IgM—in breast milk samples from patients with COVID-19.ResultsAll breast milk samples showed negative results for presence of SARS-CoV-2 RNA. We observed high intraindividual and interindividual variability in the antibody response to the receptor-binding domain of the SARS-CoV-2 spike protein for each of the three isotypes IgA, IgM and IgG. Main Protease (MPro) domain antibodies were also detected in milk. 82.9% (58 of 70) of milk samples were positive for at least one of the three antibody isotypes, with 52.9% of these positive for all three Igs. Positivity rate for IgA was relatively stable over time (65.2%–87.5%), whereas it raised continuously for IgG (from 47.8% for the first 10 days to 87.5% from day 41 up to day 206 post-PCR confirmation).ConclusionsOur study confirms the safety of breast feeding and highlights the relevance of virus-specific SARS-CoV-2 antibody transfer. This study provides crucial data to support official breastfeeding recommendations based on scientific evidence.Trial registration numberNCT04768244.

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“…Breast milk collection was performed by each women at home following the recommended procedures 16 . Then, milk was collected mainly by use of a sterile pumper in sterile bottles to normalize the collection among participants and morning collection was recommendable.…”

Section: Human Milk Collection and Processingmentioning

confidence: 99%

Anti-Sars-Cov-2 IgA And IgG In Human Milk After Vaccination Is Dependent On Vaccine Type And Previous Sars-Cov-2 Exposure: A Longitudinal Study

Selma‐Royo

Bäuerl

Mena‐Tudela

et al. 2021

Preprint

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Importance: Limited data are available on COVID-19 vaccine impact in lactating women. Objective: To evaluate the impact of different COVID-19 vaccines on specific anti-SARS-CoV-2 IgA and IgG levels in human milk. Design, Settings and Participants: In this prospective observational study in Spain, 75 lactating women from priority groups receiving vaccination against SARS-CoV-2 were included (January to April 2021). Human milk samples were collected at seven-time points. A group with confirmed SARS-CoV-2 infection (n=19) and a group of women from prepandemic time (n=13) were included. Exposure: mRNA vaccines (BNT162b2 and mRNA-1273) and adenovirus-vectored vaccine (ChAdOx1 nCoV-19). Main Outcome(s) and Measure(s): Presence of IgA and IgG against RBD SARS-CoV-2 in breast milk. Results: Seventy-five vaccinated lactating women [mean age, 34.9±3.7 years] of whom 96% were Caucasic and 92% were health care workers. A total of 417 milk samples were included and vaccine distribution was BioNTech/Pfizer (BNT162b2, n=30), Moderna (mRNA-1273, n=21), and AstraZeneca (ChAdOx1 nCoV-19, n=24). For each vaccine, 7 time points were collected from baseline up to 25 days after the 1st dose and same points were collected for mRNA vaccines 30 days after 2nd dose. A strong reactivity was observed for IgG and IgA after vaccination mainly after the 2nd dose. Presence and the persistence of specific SARS-CoV-2 antibodies in breast milk were dependent on the vaccine-type and, on previous virus exposure. High inter-variability was observed, being relevant for IgA antibodies. IgG levels were significantly higher than those observed in milk from COVID-19 women while IgA levels were lower. Women with previous COVID-19 increased the IgG levels after the 1st dose to a similar level observed in vaccinated women after the 2nd dose. Conclusions and Relevance: Breast milk from vaccinated women contains anti-SARS-CoV-2 IgA and IgG, with highest after the 2nd dose. Levels were dependent on vaccine type and previous exposure to SARS-CoV-2. Previous COVID-19 influenced the vaccine effect after a single dose, which could be especially relevant in the design of vaccination protocols . Further studies are warranted to demonstrate the potential protective role of these antibodies against COVID-19 in infants from vaccinated and infected mothers through breastfeeding.

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Best Practices for Human Milk Collection for COVID-19 Research

Cited by 25 publications

References 69 publications

Non-Coding RNAs in Human Breast Milk: A Systematic Review

Non-Coding RNAs in Human Breast Milk: A Systematic Review

SARS-CoV-2 RNA and antibody detection in breast milk from a prospective multicentre study in Spain

Anti-Sars-Cov-2 IgA And IgG In Human Milk After Vaccination Is Dependent On Vaccine Type And Previous Sars-Cov-2 Exposure: A Longitudinal Study

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