scholarly journals Improved library preparation protocols for amplicon sequencing-based noninvasive fetal genotyping for RHD-positive D antigen-negative alleles

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Asuka Hori ◽  
Hiroko Ogata-Kawata ◽  
Aiko Sasaki ◽  
Ken Takahashi ◽  
Kosuke Taniguchi ◽  
...  
Keyword(s):  
East Asian ◽  
Amplicon Sequencing ◽  
Library Preparation ◽  
Sequencing Errors ◽  
Asian Populations ◽  
Pcr Duplicates ◽  
Pcr Products ◽  
Reliability And Robustness ◽  
D Antigen ◽  
Reduced Time

Abstract Objective We aimed to simplify our fetal RHD genotyping protocol by changing the method to attach Illumina’s sequencing adaptors to PCR products from the ligation-based method to a PCR-based method, and to improve its reliability and robustness by introducing unique molecular indexes, which allow us to count the numbers of DNA fragments used as PCR templates and to minimize the effects of PCR and sequencing errors. Results Both of the newly established protocols reduced time and cost compared with our conventional protocol. Removal of PCR duplicates using UMIs reduced the frequencies of erroneously mapped sequences reads likely generated by PCR and sequencing errors. The modified protocols will help us facilitate implementing fetal RHD genotyping for East Asian populations into clinical practice.

scholarly journals Amplicon Sequencing-Based Noninvasive Fetal Genotyping for RHD-Positive D Antigen-Negative Alleles

2019 ◽  
Vol 65 (10) ◽  
pp. 1307-1316 ◽  
Author(s):  
Ken Takahashi ◽  
Ohsuke Migita ◽  
Aiko Sasaki ◽  
Michiko Nasu ◽  
Akihiro Kawashima ◽  
...  
Keyword(s):  
Pregnant Women ◽  
East Asian ◽  
Amplicon Sequencing ◽  
Sequence Difference ◽  
African Countries ◽  
Cell Free Dna ◽  
Mapping Procedure ◽  
Asian Populations ◽  
Free Dna ◽  
D Antigen

Abstract BACKGROUND To avoid hemolytic disease of the fetus and newborn resulting from maternal alloantibodies against fetal Rh antigens, anti-D immunoglobulin is routinely administered to RhD-negative pregnant women in Japan. Fetal RHD genotyping using cell-free DNA may prevent unnecessary antibody administration; however, current PCR-based methods, which detect RHD deletion, do not address the higher rates of RHD-positive D antigen-negative alleles in nonwhite populations without additional inspections. METHODS We developed an amplicon-sequencing method that could estimate the type of paternally inherited fetal RHD allele from 4 major RHD alleles in the Japanese population: the D antigen-positive allele (RHD*01, 92.9%) and 3 D antigen-negative alleles (RHD*01N.01, 6.6%; RHD*01EL.01, 0.3%; RHD*01N.04, 0.1%) using cell-free DNA obtained from the blood plasma of pregnant women. RESULTS The method correctly determined the fetal RhD type even when RhD-negative pregnant women possessed an RHD-positive D antigen-negative allele: RHD*01EL.01 or RHD*01N.04. CONCLUSIONS This method is a reliable noninvasive fetal RHD genotyping method for Japanese and other East Asian populations. The genotyping principle of amplifying 2 different regions using the same primer pair and distinguishing them by their sequence difference during the subsequent mapping procedure is also theoretically applicable to RHD-positive D antigen-negative alleles prevalent in Africans. Therefore, this method offers an opportunity to consider targeted administration of anti-D immunoglobulin to RhD-negative pregnant women in East Asian and African countries and to increase the specificity of the fetal RHD genotyping implemented nationwide in several European countries.

1188-P: Glycemic Efficacy of Sitagliptin in East Asian Populations: A Pooled Analysis

Diabetes ◽  
2019 ◽  
Vol 68 (Supplement 1) ◽  
pp. 1188-P
Author(s):  
JOAO M. CONCEICAO ◽  
CLAUDIO D. GONZALEZ ◽  
SAMUEL S. ENGEL ◽  
JONGHO AHN ◽  
SHIGERU TOKITA ◽  
...  
Keyword(s):  
East Asian ◽  
Pooled Analysis ◽  
Asian Populations

Complications of Asian Double Eyelid Surgery: Prevention and Management

2020 ◽  
Vol 36 (05) ◽  
pp. 592-601
Author(s):  
Stephanie Ming Young ◽  
Yoon-Duck Kim
Keyword(s):  
East Asian ◽  
Eyelid Surgery ◽  
Patient Dissatisfaction ◽  
Optimal Outcomes ◽  
Asian Populations ◽  
Underlying Causes ◽  
Double Eyelid

AbstractDouble eyelid surgery remains one of the most popular aesthetic surgeries, especially among East Asian populations. Complications related to double eyelid surgery can be divided into various categories: (1) patient dissatisfaction, (2) problems with the eyelid crease, (3) problems with the eyelid height, (4) suture-related complications, and (5) complications related to eyelid surgery in general. As with all eyelid surgeries, it is important to understand and appreciate the normal and abnormal function and anatomy of the Asian eyelid to reduce the risk of complications. It is also important to recognize the various complications and their underlying causes so that the surgeon can confidently revise the surgery to achieve optimal outcomes.

scholarly journals Pharmacokinetics and Pharmacodynamics of Meloxicam in East Asian Populations: The Role of Ethnicity on Drug Response

2017 ◽  
Vol 6 (12) ◽  
pp. 823-832 ◽  
Author(s):  
Takahiko Aoyama ◽  
Yoshimasa Ishida ◽  
Masato Kaneko ◽  
Aoi Miyamoto ◽  
Yoshiro Saito ◽  
...  
Keyword(s):  
Drug Response ◽  
East Asian ◽  
Pharmacokinetics And Pharmacodynamics ◽  
Asian Populations

Allozyme variation among east asian populations of the Indian rice frog Rana limnocharis (Amphibia: Anura)

1997 ◽  
Vol 25 (2) ◽  
pp. 143-159 ◽  
Author(s):  
Mamoru Toda ◽  
Mutsumi Nishida ◽  
Masafumi Matsui ◽  
Gan-Fu Wu ◽  
Hidetoshi Ota
Keyword(s):  
East Asian ◽  
Allozyme Variation ◽  
Asian Populations

Fertility Transition of the East Asian Populations.

1980 ◽  
Vol 6 (3) ◽  
pp. 501
Author(s):  
E. H. ◽  
Lee-Jay Cho ◽  
Kazumasa Kobayashi
Keyword(s):  
East Asian ◽  
Fertility Transition ◽  
Asian Populations

scholarly journals An optimized ribodepletion approach for C. elegans RNA-sequencing libraries

2021 ◽  
Author(s):  
Alec Barrett ◽  
Rebecca McWhirter ◽  
Seth R Taylor ◽  
Alexis Weinreb ◽  
David M Miller ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Noncoding Rnas ◽  
Rrna Gene ◽  
Library Preparation ◽  
C Elegans ◽  
Pcr Duplicates ◽  
Rrna Depletion ◽  
Dissociated Cells ◽  
Probe Set ◽  
Long Genes

ABSTRACTA recent and powerful technique is to obtain transcriptomes from rare cell populations, such as single neurons in C. elegans, by enriching dissociated cells using fluorescent sorting. However, these cell samples often have low yields of RNA that present challenges in library preparation. This can lead to PCR duplicates, noisy gene expression for lowly expressed genes, and other issues that limit endpoint analysis. Further, some common resources, such as sequence specific kits for removing ribosomal RNA, are not optimized for non-mammalian samples. To optimize library construction for such challenging samples, we compared two approaches for building RNAseq libraries from less than 10 nanograms of C. elegans RNA: SMARTSeq V4 (Takara), a widely used kit for selecting poly-adenylated transcripts; and SoLo Ovation (Tecan Genomics), a newly developed ribodepletion-based approach. For ribodepletion, we used a custom kit of 200 probes designed to match C. elegans rRNA gene sequences. We found that SoLo Ovation, in combination with our custom C. elegans probe set for rRNA depletion, detects an expanded set of noncoding RNAs, shows reduced noise in lowly expressed genes, and more accurately counts expression of long genes. The approach described here should be broadly useful for similar efforts to analyze transcriptomics when RNA is limiting.

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