Association of Haplotypes in Exon 4 of KLK2 Gene with Raised Serum Prostate-Specific Antigen

The standard diagnostic modalities for Prostate Cancer (PC) include serum Prostate-Specific Antigen (PSA) assay, Digital Rectal Examination (DRE), and histological examination of prostate biopsy. They are limited by low predictive potential and inability to predict which patients are at risk of developing metastatic disease. The aim of this study is to investigate the exon 4 of the KLK2 gene of subjects for changes in its nucleotide sequences (SNPs) and determine the correlation of these changes with serum PSA in an Igbo population of Nigeria. One hundred male subjects aged 40 years and above, who gave their consent, were used for the study. Their PSA determinations were done using ELISA technique while genetic studies were carried out using real-time PCR. tPSA, fPSA, and % fPSA of the subjects ranged between 0.8% - 18.30%, 0.10% - 1.60% and 0.0% - 0.7% respectively. Of the 100 subjects, 28 subjects had tPSA levels above 4.0 ng/ml with a mean of 7.10 (±3.30) ng/ml. Those with tPSA less than 4 ng/ml had a mean of 1.87 (±0.85) ng/m. 15 subjects showed SNPs with a mean tPSA of 6.87 (±4.82) ng/ml while the remaining 85 subjects without SNPs had a mean of 1.86 (±0.80) ng/ml. Results from direct DNA sequencing showed 11 SNPs. Ten subjects are curated in SNP database while one is uncurated. The Chi-square test showed significant association (p = 0.00) between tPSA levels and SNPs mutation (X2 = 17.35, p = 0.00). A Kruskal-Wallis test demonstrated that the positional arrangement of the SNP mutations had no effect on PSA-total or free-values (H (10) = 10.92, p = 0.28;H (10) = 10.07, p = 0.38 respectively). Two SNPs: rs6072 and rs74478031 were associated with elevated PSA levels (p < 0.05). Their presence, therefore, has the potential to serve, in conjunction with raised PSA, as biomarkers of prostate cancer in the study population.

Frequency of celiac disease and distribution of HLA-DQ2/DQ8 haplotypes among siblings of children with celiac disease

BACKGROUND Celiac disease(CD)is a multifactorial disease,but genetic factors play a major role in its etiology.It has been known that human leucocyte antigen(HLA)-DQ2/DQ8 haplotypes are one of the most important predisposing genetic factors.The risk of developing CD in first-degree relatives and especially siblings of celiac patients is quite high because of having the same HLA haplotypes.AIM To evaluate the frequency of CD and the distribution of the HLA-DQ2/DQ8 haplotypes in siblings of celiac patients.METHODS Patients with biopsy-proven CD and their siblings were included in the study;those who did not have HLA genotyping were excluded from the study.All siblings were on a gluten-containing diet.The HLA genotyping,tissue transglutaminase antibody IgA antibody test,and total IgA test were performed in all participants.RESULTS A total of 57 celiac patients and their 112 siblings were included in the study.The mean age of celiac patients and siblings were 10.30±3.87 years and 9.90±6.11 years,respectively.HLA-DQ2/DQ8 alleles were detected in 98.2%of patients with CD and 90.2%of siblings of celiac patients.HLA-DQ genotypes were present in all siblings diagnosed with CD.Tissue transglutaminase antibody IgA test was found to be positive in 16 siblings.CD was diagnosed in 12 siblings(10.7%)by intestinal biopsy.CONCLUSION The prevalence of CD was found to be 10.7%in siblings of celiac patients in our study.One-third of the siblings diagnosed with CD were asymptomatic.We detected HLA-DQ alleles in 98.2%of celiac patients and 100%in siblings diagnosed with CD.In addition,1 of the 2 siblings was diagnosed with CD 1 year later and the other 4 years later.Therefore,we suggest that siblings of celiac patients should be followed up with clinical findings as well as HLA analysis and serological examination.Since the risk of developing CD is much higher in asymptomatic siblings,we recommend that siblings should be screened for CD even if they are asymptomatic.

Genetic Analysis of Embryo Production Frequency in Wheat × Maize Cross

A DH population derived from C49S-87/01Y1-1069 was used to study the inheritance of wheat haploid embryo production frequency（EPF） in wheat × maize cross with the mixed major gene and polygene inheritance model of quantitative traits. The results showed that the EPF of wheat × maize cross was controlled by two dominant epistatic genes and polygene with gene effects of 1.95 for the first major gene, 6.69 for the second one and 2.80 for the polygene. The inheritability of major genes was as high as 72.09%, suggesting that the differences in EPF among wheat materials were mainly influenced by genotype. However, non-genetic factors were still important, especially for wheat materials with low EPF.

Genetic dissection and validation of a major QTL for grain weight on chromosome 3B in bread wheat(Triticum aestivum L.)

Grain weight is one of the key components of wheat(Triticum aestivum L.)yield.Genetic manipulation of grain weight is an efficient approach for improving yield potential in breeding programs.A recombinant inbred line(RIL)population derived from a cross between W7268 and Chuanyu 12(CY12)was employed to detect quantitative trait loci(QTLs)for thousand-grain weight(TGW),grain length(GL),grain width(GW),and the ratio of grain length to width(GLW)in six environments.Seven major QTLs,QGl.cib-2D,QGw.cib-2D,QGw.cib-3B,QGw.cib-4B.1,QGlw.cib-2D.1,QTgw.cib-2D.1 and QTgw.cib-3B.1,were consistently identified in at least four environments and the best linear unbiased estimation(BLUE)datasets,and they explained 2.61 to 34.85%of the phenotypic variance.Significant interactions were detected between the two major TGW QTLs and three major GW loci.In addition,QTgw.cib-3B.1 and QGw.cib-3B were co-located,and the improved TGW at this locus was contributed by GW.Unlike other loci,QTgw.cib-3B.1/QGw.cib-3B had no effect on grain number per spike(GNS).They were further validated in advanced lines using Kompetitive Allele Specific PCR(KASP)markers,and a comparison analysis indicated that QTgw.cib-3B.1/QGw.cib-3B is likely a novel locus.Six haplotypes were identified in the region of this QTL and their distribution frequencies varied between the landraces and cultivars.According to gene annotation,spatial expression patterns,ortholog analysis and sequence variation,the candidate gene of QTgw.cib-3B.1/QGw.cib-3B was predicted.Collectively,the major QTLs and KASP markers reported here provide valuable information for elucidating the genetic architecture of grain weight and for molecular marker-assisted breeding in grain yield improvement.

Morphophysiological Diversity and Haplotype Analysis of Saltol QTL Region in Diverse Rice Landraces for Salinity Tolerance

Rice is sensitive to salinity stress at both the seedling and reproductive stages.The present study used 145 rice genotypes comprising of 100 landraces and 45 advanced breeding lines collected from different regions of India.These genotypes were evaluated in hydroponics under control[electrical conductivity(ECe)~1.2 dS/m]and saline(ECe~10.0 dS/m)environments along with susceptible(IR29)and tolerant(FL478)checks.The stress susceptibility index for eight morphophysiological traits was estimated.Analysis of variance showed significant differences among the genotypes for all the parameters studied in control,stress and relative stress conditions.We identified 3 landraces(Kuttimanja,Tulasimog and IET-13713I)as tolerant and 14 lines as moderately tolerant to salt stress.Strong correlations in the morphological(root and shoot lengths)and physiological traits(shoot Na^(+),Ca^(2+)and Mg^(2+)contents,and Na^(+)/K^(+)ratio)were observed under all the conditions.The hierarchical cluster analysis grouped the genotypes into five clusters,among which cluster Ⅱ comprised salt-tolerant lines.Haplotyping of Saltol region using 11 simple sequence repeat markers on 17 saline tolerant and moderately tolerant lines was conducted.Markers AP3206F,RM10793 and RM3412b,located close to SKC1 gene(11.23‒12.55 Mb),displayed diverse allelic variations and they were not related to the FL478 type.In this region,tolerant lines like Kuttimanja,IET-13713I and Tulasimog have new alleles.As a result,these lines may be suitable candidates for novel genomic regions governing rice salinity tolerance.Salt-tolerance ability of Kuttimanja,Tulasimog and IET-13713I was validated in two years in three salinity stress environments.These promising lines can be used in breeding programs to broaden the genetic base of salinity tolerance in rice,and it may help to dissect key genomic regions responsible for salinity tolerance.

Multi-genome evolutionary study of the ABC1 gene family and identification of the pleiotropic effects of OsABC1-13 in rice development

In four rice genomes,85 ABC1-family genes were identified by comparative genomics,evolution,genetics,and physiology.One,OsABC1-13,was shown by knockdown and knockout experiments to affect plant height,grain size,and photosynthetic capability.

Haplotypes at the 5′-Flanking Region of Peroxisome Proliferator-Activated Receptor γ Gene and Their Association with the Growth and Body Composition Traits in Chickens

Peroxisome proliferator-activated receptor g（PPARg） is an important regulator of chicken preadipocyte proliferation and differentiation.In this study,polymorphisms were detected by DNA sequencing,PCR-RFLP and some other methods and three polymorphisms（g.-1784_-1768del17,c.-1241GA and c.-75GA） were found in the 5＇ flanking region of PPARg gene.Growth and body composition traits were measured in the 8th-10th generation populations of the Northeast Agricultural University broiler lines were divergently selected for abdominal fat content.Polymorphisms among individuals were screened in the above populations.The haplotype-based association analysis on growth and body composition traits was carried out.The association analysis showed that haplotypes based on three polymorphisms at 5＇ flanking region of PPARg gene were significantly associated with abdominal fat weight（AFW）,abdominal fat percentage（AFP,AFW/BW7）,liver weight（LW）,liver weight percentage（LFP,LW/BW7）,shank length（ShL）,femur weight（FeW）,keel length（KeL）,and metatarsus circle（MeC）（P0.05） and suggestive significantly associated with pectoralis major weight（PMaW）,pectoralis minor weight（PMiW）,pectoralis minor weight percentage（PMiWP,PMiW/BW7）,and metatarsus length（MeL）（P0.2）.The least square analysis showed that the birds with BGA haplotype had significantly higher AFW and AFP than the birds with other haplotypes（P0.05）.The birds with AAG haplotype had significantly higher LW and LW/BW than the birds with other haplotypes（P0.05）.The birds with AAG haplotype had significantly higher PMiW and PMiW/BW than the birds with other haplotypes（P0.05）.The birds with AAG haplotype had significantly higher ShL,FeW,MeL,MeC and KeL than the birds with AGG haplotypes（P0.05）.The results in this study revealed that QTL affecting fatness traits may exist in 5＇ flanking region of PPARg gene in chickens and PPARg gene might be one of the genes having important influences on the growth and bone traits in chickens.

Changes in plasma calcitonin gene-related peptide and serum neuron specific enolase in rats with acute cerebral ischemia after low-frequency electrical stimulation with different waveforms and intensities

Following acute cerebral ischemia in rats, plasma calcitonin gene-related peptide decreased and the level of serum neuron specific enolase and the volume of the infarction increased. Square-wave and triangular-wave electrical stimulation with low or high intensities could increase the plasma calcitonin gene-related peptide, decrease the serum neuron specific enolase and reduce the infarction volume in the brain in rats with cerebral ischemia. There was no significant difference between different wave forms and intensities. The experimental findings indicate that low-frequency electrical stimulation with varying waveforms and intensities can treat acute cerebral ischemia in rats.

A Study on the Association Between Siglec-1 Gene Polymorphism and Susceptibility in Patients with Chronic Obstructive Pulmonary Disease in Luohe Area

Objective:To analyze the association between Siglec-1 gene polymorphism and susceptibility to chronic obstructive pulmonary disease(COPD)in the population of the Luohe area.Methods:A case-control study(150 COPD patients and 150 healthy controls)was conducted to analyze the Siglec-1 allele in two groups of individuals using single nucleotide polymorphism(SNP)high-throughput detection technology,and the frequencies of each allele were compared.Results:The frequency of rs611847 heterozygous A/G genotype in COPD patients was significantly lower in females than in healthy controls(OR=0.282,95%CI=0.085-0.938,P=0.039);among smokers,the frequency of rs3859664 and rs6084444 genotypes in COPD patients was significantly higher than that in the healthy control group(OR=2.028,95%CI=1.111-3.704,P=0.021;OR=1.836,95%CI=1.033-3.262,P=0.038).Conclusion:Among the COPD population in the Luohe area,there is a significant correlation between the genotypes of three SNPs loci,rs3859664,rs6084444,and rs611847 and susceptibility to COPD in different subgroups of the population.The rs3859664 A/G-A/A and rs6084444 A/G-G/G genotypes can increase the risk of COPD in smokers;the rs611847 heterozygous A/G genotype can reduce the risk of COPD in both female and smoking populations.

ANALYZING HLA HAPLOTYPE OF THE LOCI HLA-A,-B, AND-DRB1 IN MONGOLIA ETHNIC GROUP

Objective To investigate HLA-A,-B and -DRB1 allele and HLA-A-B-DRB1 haplotype frequencies in Mongolia ethnic group. Methods HLA-A, -B, -DRB1 allele and haplotype in the Mongolia ethnic group were investigated based on 93 individuals by PCR- sequence-based typing (SBT) method. Results Twenty-one alleles were detected for HLA-A, 44 for HLA-B, and 26 for HLA-DRB1. The most frequent alleles were HLA-A*2402(0.2097), HLA-B*1302(0.0699), and HLA-DRB1*0701(0.1237). The most common HLA-A-B-DRB1 haplotype were A*3001-B*1302-DRB1*0701, A*0101-B*3701-DRB1*1001, followed by the A*0201-B*4601-DRB1*0901, A*2402-B*4801-DRB1*1101, A*2402-B*5201-DRB1*1501, A*3201-B*3503-DRB1*1301, and A*3303-B*5801-DRB1*0301, which were also presented in Chinese populations. Conclusion The data can be used in forensic and paternity tests to estimate the frequency of a DNA profile or anthropologic research. The characteristics of the distribution of HLA alleles revealed that Mongolia ethnic group is characterized by northern Mongolian Chinese.

Glutamate decarboxylase 1 gene polymorphisms are associated with respiratory symptoms in panic disorder

BACKGROUND Genetic factors play an important role in the pathogenesis of panic disorder(PD).However,the effect of genetic variants on PD remains controversial.AIM To evaluate the associations between glutamate decarboxylase 1(GAD1)gene polymorphisms and PD risk and assess the effect of GAD1 gene polymorphisms on the severity of clinical symptoms in PD.METHODS We recruited 230 PD patients and 224 healthy controls in this study.All participants were assessed for anxiety and panic symptom severity using the Hamilton Anxiety Rating Scale(HAM-A)and Panic Disorder Severity Scale(PDSS).GAD1 gene polymorphisms(rs1978340 and rs3749034)were genotyped and assessed for allele frequencies.RESULTS There were no significant differences between cases and controls in the genotype distributions or allele frequencies of GAD1(rs1978340 and rs3749034).In addition,the effect of GAD1(rs1978340 and rs3749034)on PD severity was not significant.However,regarding respiratory symptoms,patients with the GAD1 rs1978340 A/A genotype had significantly higher scores than those with the A/G or G/G genotype.CONCLUSION Here,we showed that the A/A genotype of GAD1 rs1978340 was associated with increased severity of respiratory symptoms in patients with PD.

小麦耐盐基因TaHKT1的新单倍型及其耐盐性评价

本研究利用0.5%盐胁迫对275份小麦品种(系)进行芽期耐盐性鉴定,测定发芽率、芽长、根长、根数、芽鲜质量、根鲜质量及根冠比等指标。利用主成分分析法将7个指标转化为3个主成分,累积贡献率为74.35%。通过主成分贡献率和隶属函数分析进一步将3个主成分简化为综合评价值(D)。根据D值的大小,利用聚类分析法将275份小麦品种(系)划分为高耐盐、中耐盐、耐盐、盐敏感和盐高敏感共5个等级,筛选出高耐盐种质11份、中耐盐107份、耐盐60份,累计占比64.7%。通过对13个小麦品种(系)进行TaHKT1基因序列分析,发现T∶T新单倍型与耐盐性相关。利用KASP标记对275份小麦品种(系)进行检测,有98份小麦品种(系)为新单倍型,占35.64%;新单倍型小麦品种(系)在各省份中所占比例不同。将小麦品种(系)的TaHKT1基因T∶T单倍型与耐盐性D值进行关联分析,结果表明,TaHKT1基因T∶T单倍型与小麦芽期耐盐性差异达极显著水平(p<0.01)。

甘肃省小麦品种(系)矮秆基因检测及分布规律

地方种是小麦育种的重要种质资源,为了解矮秆基因在地方种中的分布,本研究检测了甘肃省地方种矮秆基因等位变异类型及其在不同麦区的分布频率。结果表明:(1)地方种Rht-B1b和Rht-D1b的频率极低;41.4%的地方种携带Rht8,且春麦区高于冬麦区;46.7%的地方种含Rht24b,春麦区低于冬麦区。Ppd-D1a的频率仅17.8%,且春麦区低于冬麦区。另外,仅检测到Rht-D1b/Rht8、Rht-D1b/Rht24b和Rht8/Rht24b 3种组合,频率分别为0.2%、0.5%和12.8%。(2)地方种携带的矮秆基因及其组合分布频率低于育成种,且差异较大。不同来源育成品种携带的优势矮秆等位变异和频率不同,清水试验站的品种以Rht-D1b、Rht8和Rht24b为主,黄羊试验站的品种以Rht-B1b、Rht-D1b、Rht8和Rht24b为主,甘谷试验站的品种以Rht8和Rht24b为主。清水和黄羊试验站的品种秆矮、丰产性好,可在河西、沿黄灌区、陇南、陇东的小麦育种中应用;甘谷试验站的品种茎秆高,抗病性突出,可应用于定西、天水、陇南和陇东等旱地小麦的抗病改良。(3)基于分子标记检测结果,筛选出15份地方种和31份育成种,以上材料均携带2个及以上降秆基因(包括矮秆基因或Ppd-D1a),可为甘肃不同麦区小麦矮秆育种提供亲本材料。

2021年广西稻瘟病菌致病性分化及其无毒基因分析

【目的】探究广西稻瘟病菌的致病力、优势种群和优势生理小种,分析其无毒基因,了解其生理小种及无毒基因组成与分布情况,为水稻抗性育种和品种推广提供参考。【方法】使用7个我国统一鉴别品种和26份已知抗病基因的近等基因系,采用室内苗期人工喷雾接种方法对2021年从广西南部(桂南)、中部(桂中)、北部(桂北)和高寒山区4个不同生态稻作区分离得到的128株稻瘟病单孢菌株进行致病性测定。【结果】60.16%供试稻瘟病菌菌株表现出强致病力,128株稻瘟病菌株被划分为7个种群26个生理小种,ZB群为优势种群,出现频率为69.53%,优势生理小种为ZB_(13)和ZB_9,出现频率分别为25.00%、14.84%。供试菌株对26个抗病基因的毒力频率为28.12%~100.00%,其中,供试病菌对Pik、Pikm、Pi1、Pi9基因的毒力频率较低,分别为28.12%、28.91%、35.94%、37.50%。供试的广西稻瘟病菌含有与测试抗病基因相对应的无毒基因,其中15个无毒基因在桂南、桂中、桂北和高寒山区4个不同稻作区均有分布,无毒基因Avr-Pia(1)、Avr-Pia(2)、Avr-Pii、Avr-Pik^(s)、Avr-Pib、Avr-Pit、Avr-Pish(2)、Avr-Pi3(t)、Avr-Pi5(t)、Avr-Pi12(t)、Avr-Pi19(t)出现频率均低于20.00%。携带有5、6、7、8、10个无毒基因组合的菌株较多,其在供试菌株中占比分别为19.53%、11.72%、11.72%、15.63%、10.16%。【结论】2021年广西稻瘟病菌致病力强,优势种群为ZB群,优势生理小种为ZB_(13)和ZB_9,无毒基因Avr-Pia(1)、Avr-Pia(2)、Avr-Pii、Avr-Pik^(s)、Avr-Pib、Avr-Pit、Avr-Pish(2)、Avr-Pi3(t)、Avr-Pi5(t)、Avr-Pi12(t)和Avr-Pi19(t)出现频率较低,在水稻抗病育种与品种布局中,与之对应的抗病基因应慎用。

贵阳地区血小板捐献者HPA-1~6/10/15/21和HLA-A/B基因多态性研究

目的研究贵阳地区机采血小板捐献者人类血小板抗原HPA-1~6/10/15/21及人类白细胞抗原HLA-A、B基因分布及多态性。方法采用实时荧光定量PCR法(qPCR)对287名贵阳地区机采血小板捐献者进行HPA-1~6/10/15/21和HLA-A、B基因分型,列举aa\ab\bb基因及HLA-A、B等位基因的分布情况、计算aa\ab\bb基因及HLA-A、B等位基因型频率。结果287名血小板捐献者HPA-1~6/10/15/21系统中,HPA-4和HPA-10的基因型均为aa型,不具有多态性;HPA-1,2,5,6和21主要以aa型为主;仅在HPA-3和HPA-15中检出bb型;杂合度最高的是HPA-15,HPA3杂合度居次;HLA-A位点检出14个等位基因,频率最高的3个是A*02(0.36)、A*11(0.33)和A*24(0.162;HLA-B位点检出23个等位基因,频率最高的4个是B*46(0.19)、B*15(0.149、B*40(0.14)和B*13(0.13)。结论贵阳地区机采血小板捐献者HPA-1~6/10/15/21和HLA-A、B基因存在多态性,需建立该地HPA/HLA基因分型血小板供者库服务于临床。

基于高频组合片段-基因表达式编程算法的轨道交通地面沉降预测模型

[目的]地面沉降预测和控制是轨道交通盾构法隧道施工中最为关注的问题之一。为了解决现有地面沉降预测和控制中存在的模型表达过于复杂且缺乏解释性的问题,需要一种既简洁清晰,又能够描述复杂问题的可解释模型,GEP(基因表达式编程)算法提供了这种可能性,因此需对基于HFS(高频组合片段)-GEP算法的轨道交通地面沉降预测模型进行深入研究。[方法]以杭绍城际铁路某区段盾构隧道工程为依托,选取盾构施工过程中的土舱压力、刀盘扭矩、刀盘转速、推进速度、总推力、隧道埋深及盾尾注浆量等参数作为关键输入型施工参数,地面沉降作为输出型施工参数,通过备选公式集筛选以及HFS选取,建立基于HFS-GEP算法的轨道交通地面沉降预测模型。利用该模型对第180环—第210环区段的关键施工参数进行优化调整,分析盾构施工参数变化对地面最终沉降的影响效果。[结果及结论]基于HFS-GEP算法的地面沉降预测模型可以反映盾构施工参数与地面最终沉降的显式关系;相较于传统GEP算法的地面沉降预测模型,该模型准确度更高,结构更为简洁,且收敛速度更快。通过对盾构关键施工参数进行优化调整,该模型可将第180环—第210环区段的最终沉降量控制在10 mm以内。

Genetic Structure of the Oriental River Prawn (Macrobrachium nipponense) from the Yangtze and Lancang Rivers, Inferred from COI Gene Sequence

This study analyzed nueleotide sequences from the mitochondrial eytochrome oxidase submit （COI） gene region （450 bp） to investigate the genetic structure of the oriental river prawn （ Macrobrachium nipponense ） among nine populations from the Yangtze and Lancang Rivers. A total of 79 individuals were collected for this work. Eighty-nine nucleotides were found to be variable, resulting in 46 haplotypes. Among the nine populations, the population from Kunming shows the greatest level of variability （h = 1.000, π = 0.028）, whereas the population from Cbongqing exhibits the lowest level of variability （h = 0.700,π = 0.008）. Analysis of molecular variance suggested that of the total genetic diversity, 9.66% was attributable to inter-population diversity and the remainder （90.34%） to differences within populations. A molecular phylogenetic tree constructed using the Neighbor-joining （N J） method showed that the 46 haplotypes were assigned to two clades associated with geographic regions. These results provide basic information for the conservation and sustainable exploitation of this species.

血小板HPA-3,HPA-15基因分型微滴式数字PCR检测体系的构建

目的建立血小板HPA-3,HPA-15基因分型的微滴式数字PCR(ddPCR)高灵敏检测方法,并初步探索应用于孕妇外周血胎儿游离DNA HPA抗原相容性检测的可行性。方法针对HPA-3,HPA-15的SNP突变位点,设计特异性引物及MGB探针,优化ddPCR退火温度及引物浓度等扩增条件,建立最佳反应体系,明确检验程序。对该检测方法进行方法学性能评估包括特异性、灵敏度、重复性和稳定性。利用ddPCR技术对2022年6月至2023年6月67例临床血液标本进行检测,将等位基因分型结果与基因测序结果比较,并对52例母体外周血胎儿游离DNA HPA抗原进行检测。结果检测血小板HPA-3,HPA-15的ddPCR方法,引物及探针特异性良好,HPA-3,HPA-15的最佳退火温度分别为:61.6℃,60.2℃;体系最佳引物浓度分别为:900 nM,700 nM;探针终浓度均为250 nM。拷贝数定量检测范围为:2~20000 copies,检测下限为0.1 copies/μL且线性良好。在低拷贝数标本中,HPA-3及HPA-15实际检测值的批内及批间变异系数(CV)均<5%。对67份人血液标本DNA的HPA-3,HPA-15基因型检测,结果与基因测序结果完全一致。应用于胎母血小板HPA-3,HPA-15基因型检测结果符合预期。结论本研究构建的HPA-3,HPA-15 ddPCR检测体系准确性高,重复性及稳定性较好,灵敏度高,可应用于临床血小板HPA-3,HPA-15基因型供者库的建立、基因配型及胎母血小板相容性检测等。