Distribution of Human Cytomegalovirus Glycoprotein H Genotypes in Different Samples from Chinese Children

2021 ◽  
Author(s):  
Wei Li ◽  
Lu-Yan Chen ◽  
Ran Tao ◽  
Shi-Qiang Shang
Keyword(s):  
Breast Milk ◽  
Human Cytomegalovirus ◽  
Throat Swab ◽  
Quantitative Polymerase Chain Reaction ◽  
Age Groups ◽  
Urine Samples ◽  
Serum Samples ◽  
Predominant Genotype ◽  
Milk Samples ◽  
Glycoprotein H

Abstract Objective This study aimed to investigate characteristics of human cytomegalovirus (HCMV) glycoprotein H (gH) genotypes in urine, throat swab, and serum from children and breast milk from children's mothers. Methods Fresh urine samples, throat swabs, or serum samples from children and breast milk samples from children's mothers were collected for HCMV DNA detection. The positive samples of HCMV DNA were further detected by fluorescent quantitative polymerase chain reaction (PCR) with gH typing. Results Of 1,703 HCMV DNA-positive samples, the highest proportion (83.3%, 85/102) of children aged between 21 days and 3 months was detected positive in breast milk samples (p = 0.002), and the highest proportion (70.5%, 110/156) of children aged above 3 months was detected positive in throat swab samples (p = 0.002). HCMV in throat swab specimens is mainly high copy (p < 0.0001), and low-copy HCMV is prevalent in breast specimens (p < 0.0001). Among them, 1,059 samples were identified as gH1 genotype, 530 samples were gH2, and 114 samples were coinfection (gH1/2). There had the highest gH2 rates (32.3%) and lowest gH1 (61.0%) rates in urine samples (p = 0.041), whereas the highest gH1 rates (71.6%) and lowest gH2 rates (19.6%) were found in breast milk samples (p = 0.032). Concerning age groups, patients aged between 21 days and 3 months had the highest gH1 proportion (p = 0.017), while patients aged above 3 months had the highest gH1 and gH2 HCMV coinfection proportion (p = 0.002). Among 43 pairs of maternal and child samples corresponding to positive samples, gH genotype of 35 pairs of samples was consistent with a rate of 81.4%. Conclusion gH1 is the predominant genotype of HCMV in each kind of sample in China. However, the distribution of the HCMV gH genotype is different among different samples.

scholarly journals COVID-19 Vaccine mRNABNT162b2 Elicits Human Antibody Response in Milk of Breastfeeding Women

Vaccines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 785
Author(s):  
Maurizio Guida ◽  
Daniela Terracciano ◽  
Michele Cennamo ◽  
Federica Aiello ◽  
Evelina La Civita ◽  
...  
Keyword(s):  
Breast Milk ◽  
Antibody Response ◽  
Human Milk ◽  
Human Antibody ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Low Concentration ◽  
Milk Samples ◽  
Milk Antibodies ◽  
Roche Diagnostics

Objective: The objective of this research is to demonstrate the release of SARS-CoV-2 Spike (S) antibodies in human milk samples obtained by patients who have been vaccinated with mRNABNT162b2 vaccine. Methods: Milk and serum samples were collected in 10 volunteers 20 days after the first dose and 7 seven days after the second dose of the mRNABNT162b2 vaccine. Anti-SARS-CoV-2 S antibodies were measured by the Elecsys® Anti-SARS-CoV-2 S ECLIA assay (Roche Diagnostics AG, Rotkreuz, Switzerland), a quantitative electrochemiluminescence immunometric method. Results: At first sample, anti-SARS-CoV-2 S antibodies were detected in all serum samples (103.9 ± 54.9 U/mL) and only in two (40%) milk samples with a low concentration (1.2 ± 0.3 U/mL). At the second sample, collected 7 days after the second dose, anti-SARS-CoV-2 S antibodies were detected in all serum samples (3875.7 ± 3504.6 UI/mL) and in all milk samples (41.5 ± 47.5 UI/mL). No correlation was found between the level of serum and milk antibodies; the milk antibodies/serum antibodies ratio was on average 2% (range: 0.2–8.4%). Conclusion: We demonstrated a release of anti-SARS-CoV-2 S antibodies in the breast milk of women vaccinated with mRNABNT162b2. Vaccinating breastfeeding women could be a strategy to protect their infants from COVID-19 infection.

scholarly journals Human Cytomegalovirus Genome Diversity in Longitudinally Collected Breast Milk Samples

2021 ◽  
Vol 11 ◽  
Author(s):  
Jasper Götting ◽  
Katrin Lazar ◽  
Nicolás M. Suárez ◽  
Lars Steinbrück ◽  
Tabea Rabe ◽  
...  
Keyword(s):  
Breast Milk ◽  
Human Cytomegalovirus ◽  
High Throughput Sequencing ◽  
De Novo ◽  
Severe Disease ◽  
Viral Population ◽  
Preterm Neonates ◽  
Genotype Distribution ◽  
Milk Samples ◽  
Wide Range

Reactivation and shedding of human cytomegalovirus (HCMV) in breast milk during lactation is highly frequent in HCMV-seropositive mothers. This represents a key transmission route for postnatal HCMV infection and can lead to severe disease in preterm neonates. Little is known about HCMV strain composition or longitudinal intrahost viral population dynamics in breast milk from immunocompetent women. We performed HCMV-specific target enrichment and high-throughput sequencing of 38 breast milk samples obtained in Germany between days 10 and 60 postpartum from 15 mothers with HCMV DNA lactia, and assembled HCMV consensus sequences de novo. The genotype distribution and number of HCMV strains present in each sample were determined by quantifying genotype-specific sequence motifs in 12 hypervariable viral genes, revealing a wide range of genotypes (82/109) for these genes in the cohort and a unique, longitudinally stable strain composition in each mother. Reactivation of up to three distinct HCMV strains was detected in 8/15 of mothers, indicating that a representative subset of the woman’s HCMV reservoir might be locally reactivated early during lactation. As described previously, nucleotide diversity of samples with multiple strains was much higher than that of samples with single strains. Breast milk as a main source of postnatal mother-to-infant transmission may serve as a repository for viral diversity and thus play an essential role in the natural epidemiology of HCMV.

scholarly journals Detection and Prevalence of Canine Leptospirosis from Navsari District of South Gujarat, India

2020 ◽  
pp. 103-110
Author(s):  
Dhruv Desai ◽  
Pushpa Makwana ◽  
Jayesh Solanki ◽  
Irshadullakhan Kalyani ◽  
Dharmesh Patel ◽  
...  
Keyword(s):  
Solid Phase ◽  
Age Groups ◽  
Dark Field ◽  
Urine Samples ◽  
Leptospira Interrogans ◽  
Age Group ◽  
Serum Samples ◽  
Percent Positivity ◽  
Significant Difference ◽  
Dot Elisa

Aims: Leptospirosis is endemically prevalent in coastal area of south Gujarat. Generally, pets are vaccinated against Leptospira interrogans but whosoever left unvaccinated, are prone to infection. The present study was conducted to detect canine leptospirosis and its prevalence from south Gujarat. Place and Duration of Study: A total of 46 serum samples and 33 urine samples were collected from 56 dogs suspected of having leptospira infection. The study was done from September 2019 to May 2020. Methodology: Serum samples were diagnosed by IgG based ImmunoComb solid phase dot ELISA test for antibody against genus specific Leptospira interrogans. Urine samples were visualized under dark field microscopy (DFM) following laboratory manual on leptospirosis of WHO. Sex and age of dogs were noted to find out the percent positivity and prevalence. Chi square test was applied to find out any significant difference among the age groups. Results: Out of 46 serum samples, 17 (36.95%) found positive by dot ELISA and out of 33 urine sample, 13 (39.39%) found positive by DFM. Sex wise prevalence found 42.10 percent (16/38) in Male while 55.55 percent (10/18) in female dogs. Age group wise prevalence was recorded 63.63 percent (07/11) in 1- ≤ 3 years group. 27.77 per cent (05/18) in > 3- ≤ 6 years group whereas, 51.85 percent (14/27) in > 6- ≤ 12 years group. Conclusion: Detection and prevalence of canine leptospirosis were studied first time from Gujarat. Overall 46.42 percent (26/56) dogs were diagnosed positive for canine leptospirosis. Dogs having age group of 1- ≤ 3 years found more susceptible. Prevalence was found more in female than male dogs. 

Impact of mode of delivery on the milk microbiota composition of healthy women

2015 ◽  
Vol 7 (1) ◽  
pp. 54-60 ◽  
Author(s):  
R. Cabrera-Rubio ◽  
L. Mira-Pascual ◽  
A. Mira ◽  
M. C. Collado
Keyword(s):  
Breast Milk ◽  
Biological Effects ◽  
Quantitative Polymerase Chain Reaction ◽  
Mode Of Delivery ◽  
Principal Coordinates Analysis ◽  
Microbiome Composition ◽  
Milk Samples ◽  
Principal Coordinates ◽  
Vaginal Deliveries ◽  
The Impact

Breast milk constitutes one of the most important sources of postnatal microbes. However, the influence of perinatal factors on the milk microbiome is still poorly understood. The aim of our study was to assess the impact of mode of delivery on the microbiome composition and diversity present in breast milk of healthy mothers. Mature milk samples (n=10) were taken from mothers after 1 month of exclusively breastfeeding. Microbiomes from milk samples were analyzed with 16S ribosomal RNA gene pyrosequencing and targeted quantitative polymerase chain reaction (PCR). Despite inter-individual variability in bacterial composition, The Principal Coordinates Analysis clearly separated milk microbiome from mothers with vaginal delivery (n=6) from those who undergo C-section (n=4). In addition, higher bacterial diversity and richness was found in milk samples from vaginal deliveries. Quantitative PCR data showed that higher levels of Bifidobacterium spp. were related significantly to lower levels of Staphylococcus spp. Despite the low sample size, our data suggest that mode of delivery has an important impact on milk microbiome composition. Further studies with larger sample sizes are needed to confirm these results and to understand the biological effects of C-section associated microbes on infant’s health.

scholarly journals Natalizumab during pregnancy and lactation

2017 ◽  
Vol 24 (12) ◽  
pp. 1627-1634 ◽  
Author(s):  
Undine Proschmann ◽  
Katja Thomas ◽  
Sandra Thiel ◽  
Kerstin Hellwig ◽  
Tjalf Ziemssen
Keyword(s):  
Breast Milk ◽  
Cell Sorting ◽  
Hl60 Cell ◽  
Facs Analysis ◽  
Serum Samples ◽  
Milk Samples ◽  
Late Antigen ◽  
Pregnancy And Lactation ◽  
Potential Risks ◽  
Exposure During Pregnancy

Background: Managing medication during pregnancy and lactation in multiple sclerosis (MS) patients needs to balance potential risks to the newborn with the substantial risks of ongoing disease activity. Objective: To evaluate the potential transfer of natalizumab (NAT) into breast milk and into the serum of newborn babies in women who continued NAT treatment during pregnancy and lactation. Methods: Serum samples of 11 mother–infant pairs and mother milk samples of a further 4 women were analyzed for free NAT using a HL60 cell-based fluorescence-activated cell sorting (FACS) assay. Two mother–baby pairs were analyzed for cell-bound NAT, very-late-antigen (VLA)-4 expression, and saturation with NAT on immune cells by FACS analysis. Results: In the majority of the mother–infant serum pairs (6/11) and in all breast milk samples, free NAT was detectable. Cell-bound NAT was measurable in both mother–baby pairs with significant higher levels in babies. VLA-4 expression seems to be higher in newborns. Saturation with NAT was comparable between newborns and mothers. Conclusion: NAT can pass placental barrier before delivery and into breast milk. Measurable NAT on neonatal lymphocytes may have functional impact. Further investigations are needed to clarify safety and risk of NAT exposure during pregnancy and lactation.

scholarly journals Multiple-Strain Infections of Human Cytomegalovirus With High Genomic Diversity Are Common in Breast Milk From Human Immunodeficiency Virus–Infected Women in Zambia

2019 ◽  
Vol 220 (5) ◽  
pp. 792-801 ◽  
Author(s):  
Nicolás M Suárez ◽  
Kunda G Musonda ◽  
Eric Escriva ◽  
Margaret Njenga ◽  
Anthony Agbueze ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Breast Milk ◽  
Human Cytomegalovirus ◽  
Developed Countries ◽  
Genomic Diversity ◽  
Viral Loads ◽  
Strain Diversity ◽  
Milk Samples ◽  
Immunodeficiency Virus ◽  
Hiv Negative

Abstract Background In developed countries, human cytomegalovirus (HCMV) is a major pathogen in congenitally infected and immunocompromised individuals, where multiple-strain infection appears linked to disease severity. The situation is less documented in developing countries. In Zambia, breast milk is a key route for transmitting HCMV and carries higher viral loads in human immunodeficiency virus (HIV)–infected women. We investigated HCMV strain diversity. Methods High-throughput sequence datasets were generated from 28 HCMV-positive breast milk samples donated by 22 mothers (15 HIV-infected and 7 HIV-negative) at 4–16 weeks postpartum, then analyzed by genome assembly and novel motif-based genotyping in 12 hypervariable HCMV genes. Results Among the 20 samples from 14 donors (13 HIV-infected and one HIV-negative) who yielded data meeting quality thresholds, 89 of the possible 109 genotypes were detected, and multiple-strain infections involving up to 5 strains per person were apparent in 9 HIV-infected women. Strain diversity was extensive among individuals but conserved compartmentally and longitudinally within them. Genotypic linkage was maintained within hypervariable UL73/UL74 and RL12/RL13/UL1 loci for virus entry and immunomodulation, but not between genes more distant from each other. Conclusions Breast milk from HIV-infected women contains multiple HCMV strains of high genotypic complexity and thus constitutes a major source for transmitting viral diversity.

scholarly journals Multiple-Strain Infections of Human Cytomegalovirus with High Genomic Diversity are Common In Breast Milk from HIV-Positive Women in Zambia

2018 ◽  
Author(s):  
Nicolás M. Suárez ◽  
Kunda G. Musonda ◽  
Eric Escriva ◽  
Margaret Njenga ◽  
Anthony Agbueze ◽  
...  
Keyword(s):  
Breast Milk ◽  
Human Cytomegalovirus ◽  
Developed Countries ◽  
Genomic Diversity ◽  
Hiv Positive ◽  
Viral Loads ◽  
Strain Diversity ◽  
Milk Samples ◽  
Hiv Positive Women ◽  
Hiv Negative

ABSTRACTBackgroundIn developed countries, human cytomegalovirus (HCMV) is a major pathogen in congenitally infected and immunocompromised individuals, in whom multiple-strain infection is linked to disease severity. The situation is less documente in developing countries. In Zambia, breast milk is a key route for transmitting HCMV and carries higher viral loads in HIV-positive women. We investigated HCMV strain diversity.MethodsHigh-throughput sequence datasets were generated from 28 HCMV-positive breast milk samples donated by 22 mothers (15 HIV-positive and seven HIV-negative) at 4 or 16 weeks (or both) postpartum and analysed by genotyping 12 hypervariable HCMV genes.ResultsAmong the 20 samples from 14 donors (13 HIV-positive and one HIV-negative) that yielded data meeting quality thresholds, 89 of the possible 109 genotypes were detected, and multiple-strain infections involving up to five strains per person were apparent in nine HIV-positive women. Strain diversity was extensive among individuals but conserved compartmentally and longitudinally within them. Genotypic linkage was maintained within the hypervariable UL73/UL74 and RL12/RL13/UL1 loci for virus-entry and immunomodulation, but not between genes more distant from each other.ConclusionsBreast milk from HIV-positive women contains multiple HCMV strains of high genotypic complexity and thus constitutes a major source for transmitting viral diversity.

scholarly journals AB1294-HPR LOW CONCENTRATIONS OF BIOLOGIC DMARDS IN BREASTMILK OF PATIENTS TREATED DURING LACTATION

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1937.1-1937
Author(s):  
A. Zbinden ◽  
K. Eriksson ◽  
F. Förger
Keyword(s):  
Breast Milk ◽  
Certolizumab Pegol ◽  
Average Concentration ◽  
Limited Information ◽  
Serum Samples ◽  
Limit Of Quantification ◽  
Biologic Dmards ◽  
Breastfed Infants ◽  
Milk Samples ◽  
Infant Dose

Background:There is very limited information about the passage of biologics into breast milk and into the peripheral blood of breastfed infants. However, based on pharmacological properties of biologic DMARDs (bDMARDs) lactation may not be discouraged in patients with chronic inflammatory rheumatic disease to treat or prevent postpartum relapses. We here report two cases treated with bDMARDs during lactation: one woman with Muckle-Wells syndrome (MWS) treated with canakinumab and one woman with microscopic polyangiitis (MPA) treated with rituximab.Objectives:To determine the level of rituximab and canakinumab in breast milk, in sera of breastfed infants as well as in sera of the mother and to calculate the average daily infant dose and the relative infant dose.Methods:Serum and milk levels of Rituximab were measured by ELISA using commercially available coating and detection antibodies. For Canakinumab an ELISA was established by coating of plates with recombinant human IL-1beta and detection of Canakinumab in samples by a polyclonal anti-human IgG coupled to HRP. In both cases separate standard curves for serum and milk were established. Serum samples and milk samples of unexposed healthy controls were used to determine the lower limit of quantification.Results:One patient with MWS received canakinumab 150 mg s.c. to treat a worsening of her disease ten days postpartum. She continued to breastfeed her child. The average concentration of canakinumab in milk samples collected on 10 consecutive days was 15.8 ng/ml. The average daily infant dose was 0.002 mg/kg/day. The relative infant dose, which refers infant to maternal exposure on a dose/weight basis, was 0.11%. There was no detectable canakinumab in the serum of the infant.One patient with MPA received rituximab 500 mg i.v. as a remission maintenance therapy four months postpartum. She continued to breastfeed her child. The average concentration of rituximab in milk samples collected on 4 consecutive days was 3.71 ng/ml. The average daily infant dose was 0.001 mg/kg/day. The relative infant dose was 0.01%. There was no detectable rituximab in the serum of the infant.Conclusion:Only minimal concentrations of canakinumab and rituximab can be detected in breastmilk. For both bDMARDs, the relative infant dose was below 1% of the maternal dose, which is considered unlikely to be of clinical concern. The lack of detectable levels of canakinumab and rituximab in the infants’ sera supports the notion of low oral bioavailability of large monoclonal antibodies. Together, the results are similar to those seen in TNF inhibitors which are regarded to be compatible with breastfeeding, yet more data are needed (1, 2, 3).References:[1]Götestam Skorpen C, Hoeltzenbein M, Tincani A, Fischer-Betz R, Elefant E, Chambers C, da Silva J, Nelson-Piercy C, Cetin I, Costedoat-Chalumeau N, Dolhain R, Förger F, Khamashta M, Ruiz-Irastorza G, Zink A, Vencovsky J, Cutolo M, Caeyers N, Zumbühl C, Østensen M. The EULAR points to consider for use of antirheumatic drugs before pregnancy, and during pregnancy and lactation. Ann Rheum Dis. 2016 May;75(5):795-810. doi: 10.1136/annrheumdis-2015-208840. Epub 2016 Feb 17.[2]Clowse ME, Förger F, Hwang C, Thorp J, Dolhain RJ, van Tubergen A, Shaughnessy L, Simpson J, Teil M, Toublanc N, Wang M, Hale TW. Minimal to no transfer of certolizumab pegol into breast milk: results from CRADLE, a prospective, postmarketing, multicentre, pharmacokinetic study. Ann Rheum Dis. 2017 Nov;76(11):1890-1896. doi: 10.1136/annrheumdis-2017-211384. Epub 2017 Aug 16.[3]Matro R, Martin CF, Wolf D, Shah SA, Mahadevan U. Exposure Concentrations of Infants Breastfed by Women Receiving Biologic Therapies for Inflammatory Bowel Diseases and Effects of Breastfeeding on Infections and Development.Gastroenterology. 2018;155(3):696–704. doi:10.1053/j.gastro.2018.05.040Disclosure of Interests:Astrid Zbinden: None declared, Klara Eriksson: None declared, Frauke Förger Grant/research support from: Unrestricted grant from UCB, Consultant of: UCB, GSK, Roche, Speakers bureau: UCB, GSK

scholarly journals Development and validation of a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of ZIKV in patient samples from Brazil

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Severino Jefferson Ribeiro da Silva ◽  
Keith Pardee ◽  
Udeni B. R. Balasuriya ◽  
Lindomar Pena
Keyword(s):  
Reverse Transcription ◽  
Rapid Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Extraction ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Serum Samples ◽  
Low Resource ◽  
Loop Mediated Isothermal Amplification ◽  
One Step

AbstractWe have previously developed and validated a one-step assay based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of the Zika virus (ZIKV) from mosquito samples. Patient diagnosis of ZIKV is currently carried out in centralized laboratories using the reverse transcription-quantitative polymerase chain reaction (RT-qPCR), which, while the gold standard molecular method, has several drawbacks for use in remote and low-resource settings, such as high cost and the need of specialized equipment. Point-of-care (POC) diagnostic platforms have the potential to overcome these limitations, especially in low-resource countries where ZIKV is endemic. With this in mind, here we optimized and validated our RT-LAMP assay for rapid detection of ZIKV from patient samples. We found that the assay detected ZIKV from diverse sample types (serum, urine, saliva, and semen) in as little as 20 min, without RNA extraction. The RT-LAMP assay was highly specific and up to 100 times more sensitive than RT-qPCR. We then validated the assay using 100 patient serum samples collected from suspected cases of arbovirus infection in the state of Pernambuco, which was at the epicenter of the last Zika epidemic. Analysis of the results, in comparison to RT-qPCR, found that the ZIKV RT-LAMP assay provided sensitivity of 100%, specificity of 93.75%, and an overall accuracy of 95.00%. Taken together, the RT-LAMP assay provides a straightforward and inexpensive alternative for the diagnosis of ZIKV from patients and has the potential to increase diagnostic capacity in ZIKV-affected areas, particularly in low and middle-income countries.

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