Molecular genotyping

分子分型
  • 文章类型: Journal Article
    当前的研究调查了肺炎支原体(M.肺炎)在上海,为上海地区肺炎支原体感染的临床治疗提供参考。
    从2017年至2019年上海儿科患者的鼻咽抽吸物中分离出临床菌株。三种抗菌类大环内酯的九种抗菌剂,氟喹诺酮类和四环素类,使用肉汤微量稀释法对肺炎支原体分离株进行了研究。通过评估23SrRNA基因和核糖体蛋白基因L4和L22的序列来分析大环内酯抗性机制。进行分子基因分型以对P1亚型和多位点可变数串联重复分析(MLVA)类型进行分类。
    根据23SrRNA基因中的A2063G突变,共有72个分离株对大环内酯类抗生素(红霉素的MIC>64mg/L)具有抗性。这些菌株对四环素和氟喹诺酮类药物敏感。P1型1(166/182,91.2%)和MLVA型4-5-7-2(165/182,90.7%)是主要亚型。MLVA类型与P1亚型相关。P1亚型和MLVA类型的分布不随时间变化。在为期三年的研究中,P12型和MLVA3-5-6-2型菌株的大环内酯耐药率增加。5基因座MLVA分型方案揭示了2019年对大环内酯耐药的MLVA3-4-5-7-2型菌株的克隆扩增。
    上海肺炎支原体的大环内酯耐药性非常高,并且在某些亚型之间不断发展。应注意大环内酯抗性基因型在该人口稠密区域内可能的克隆传播。
    UNASSIGNED: The current study investigated the recent genetic characteristics and antimicrobial susceptibility profiles of Mycoplasma pneumoniae (M. pneumoniae) in Shanghai, becoming a clinical reference for treating M. pneumoniae infection in Shanghai.
    UNASSIGNED: Clinical strains were isolated from nasopharyngeal aspirates of the pediatric patients in Shanghai from 2017 to 2019. Nine antimicrobial agents of three antimicrobial classes macrolides, fluoroquinolones and tetracyclines, against M. pneumoniae isolates were investigated using the broth microdilution method. The mechanism of macrolide resistance was analyzed by evaluating the sequences of the 23S rRNA gene and the ribosomal protein genes L4 and L22. Molecular genotyping was undergone to classify the P1 subtypes and the multi-locus variable-number tandem-repeat analysis (MLVA) types.
    UNASSIGNED: A total of 72 isolates were resistant to macrolides (MICs > 64 mg/L for erythromycin) based on the A2063G mutation in the 23S rRNA gene. These strains were susceptible to tetracyclines and fluoroquinolones. P1 type 1 (166/182, 91.2%) and MLVA type 4-5-7-2 (165/182, 90.7%) were the dominant subtypes. MLVA type was associated with the P1 subtypes. The distribution of the P1 subtypes and MLVA types did not change over time. The macrolide-resistant rate in P1 type 2 and MLVA type 3-5-6-2 strains were increased during the three-year study. The 5-loci MLVA typing scheme revealed the clonal expansion of MLVA type 3-4-5-7-2 strains which are macrolide-resistant in 2019.
    UNASSIGNED: Macrolide resistance in M. pneumoniae in Shanghai is very high and is evolving among certain subtypes. Cautions should be taken for the possible clonal spreading of macrolide-resistant genotypes within this populated region.
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  • 文章类型: Journal Article
    背景:侵袭性念珠菌病在全球范围内日益受到关注,尤其是在免疫功能低下的患者中,包括ICU患者。
    目的:由于白色念珠菌是导致念珠菌血症的主要原因,研究念珠菌血症患者白色念珠菌的演变过程非常重要。
    方法:我们分析了从不同身体部位分离的238株白色念珠菌。抗真菌药敏试验,对所有分离株进行了CAI基因座基因分型和多位点序列分型(MLST)。使用eBURST分析了仅在七个管家基因基因座之一上序列不同的总分离株之间的关系。
    结果:通过结合七个管家等位基因对238个分离株进行的多位点序列分型分析揭示了175个二倍体序列类型,其中84个是新确定的。对这些数据的eBURST分析识别了19个克隆复合物(CC)和79个单例。此外,鉴定出73种CAI基因型。血液分离株显示最大基因型(49),显性基因型为CAI17-21和CAI21-21。口腔分离株具有25种CAI基因型,显性基因型为CAI17-21和CAI21-21。由于CAI等位基因数<30的分离株显示更容易传播,CAI17-21和CAI21-21是最常见的传播。最后,CAI基因型分为6组。
    结论:这项工作揭示了从ICU念珠菌血症患者中分离出的口腔和血液菌株具有相同的显性CAI基因型。我们的数据扩展了白色念珠菌MLST数据库,并有助于了解侵袭性念珠菌病的演变和传播。
    BACKGROUND: Invasive candidiasis is a growing concern worldwide, especially in immunocompromised patients, including ICU patients.
    OBJECTIVE: As Candida albicans is the leading cause of candidaemia, it is important to investigate the evolution of C. albicans in patients with candidaemia.
    METHODS: We analysed 238 strains of C. albicans isolated from different body sites. Antifungal susceptibility testing, CAI loci genotyping and multilocus sequence typing (MLST) of all isolates were performed. The relationships among the total isolates that differed in sequence at only one of the seven housekeeping gene loci were analysed using eBURST.
    RESULTS: Multilocus sequence typing analysis in 238 isolates by combining seven housekeeping alleles revealed 175 diploid sequence types, in which 84 were newly identified. eBURST analysis for these data recognised 19 clonal complexes (CCs) and 79 singletons. Besides, seventy-three CAI genotypes were identified. Blood isolates showed maximum genotypes (49), and the dominant genotypes were CAI 17-21 and CAI 21-21. Oral isolates possessed 25 CAI genotypes, and the dominant genotypes were CAI 17-21 and CAI 21-21 as well. Since isolates with CAI allele numbers <30 showed easier transmission, CAI 17-21 and CAI 21-21 were the most frequently transmitted. Finally, the CAI genotypes were classified into six groups.
    CONCLUSIONS: This work revealed the oral and blood strains isolated from the patients with candidaemia in ICU shared the identical dominant CAI genotypes. Our data expanded the C. albicans MLST database and helped with understanding the evolution and spread of invasive candidiasis.
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  • 文章类型: Journal Article
    保持遗传完整性对于作物野生近缘(CWR)物种的原位和非原位保护至关重要。然而,作物等位基因渗入CWR物种/种群可能会改变其遗传结构和多样性,导致更具侵入性的杂草,相比之下,濒危物种的灭绝。为了确定作物野生渗入及其后果,我们研究了中国就地保护下的六个野生稻(Oryzarufipogon)种群的遗传结构和多样性。使用34个简单序列重复序列(SSR)和34个插入/缺失标记对野生稻种群和两组水稻品种(O.sativa),对应于两种类型的分子标记。共有等位基因和结构分析表明,作物野生基因渗入和混合水平可变。主坐标和聚类分析表明野生稻种群的分化,这与到栽培稻田的空间距离有关。野生稻种群和水稻品种之间的总体遗传多样性水平相当,但是在野生种群中检测到大量的野生特异性等位基因。我们根据结果得出结论,作物野生基因渗入可以大大改变CWR种群的遗传结构和关系的模式。在作物野生渗入的情况下,应采取适当措施对CWR物种进行有效的就地保护。
    Maintaining genetic integrity is essential for in situ and ex situ conservation of crop wild relative (CWR) species. However, introgression of crop alleles into CWR species/populations may change their genetic structure and diversity, resulting in more invasive weeds or, in contrast, the extinction of endangered populations. To determine crop-wild introgression and its consequences, we examined the genetic structure and diversity of six wild rice (Oryza rufipogon) populations under in situ conservation in China. Thirty-four simple sequence repeat (SSR) and 34 insertion/deletion markers were used to genotype the wild rice populations and two sets of rice cultivars (O. sativa), corresponding to the two types of molecular markers. Shared alleles and STRUCTURE analyses suggested a variable level of crop-wild introgression and admixture. Principal coordinates and cluster analyses indicated differentiation of wild rice populations, which was associated with the spatial distances to cultivated rice fields. The level of overall genetic diversity was comparable between wild rice populations and rice cultivars, but a great number of wild-specific alleles was detected in the wild populations. We conclude based on the results that crop-wild introgression can considerably alter the pattern of genetic structure and relationships of CWR populations. Appropriate measures should be taken for effective in situ conservation of CWR species under the scenario of crop-wild introgression.
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  • 文章类型: Journal Article
    Lack of a consistent and reliable genotyping system can critically impede HIV genomic research on pathogenesis, fitness, virulence, drug resistance, and genomic-based healthcare and treatment. At present, mis-genotyping, i.e., background noises in molecular genotyping, and its impact on epidemic surveillance is unknown. For the first time, we present a comprehensive assessment of HIV genotyping quality. HIV sequence data were retrieved from worldwide published records, and subjected to a systematic genotyping assessment pipeline. Results showed that mis-genotyped cases occurred at 4.6% globally, with some regional and high-risk population heterogeneities. Results also revealed a consistent mis-genotyping pattern in gp120 in all studied populations except the group of men who have sex with men. Our study also suggests novel virus diversities in the mis-genotyped cases. Finally, this study reemphasizes the importance of implementing a standardized genotyping pipeline to avoid genotyping disparity and to advance our understanding of virus evolution in various epidemiological settings.
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  • 文章类型: Journal Article
    Germplasm collections provide an extremely valuable resource for breeders and researchers. However, misclassification of accessions by species often hinders the effective use of these collections. We propose that use of high-throughput genotyping tools can provide a fast, efficient and cost-effective way of confirming species in germplasm collections, as well as providing valuable genetic diversity data. We genotyped 180 Brassicaceae samples sourced from the Australian Grains Genebank across the recently released Illumina Infinium Brassica 60K SNP array. Of these, 76 were provided on the basis of suspected misclassification and another 104 were sourced independently from the germplasm collection. Presence of the A- and C-genomes combined with principle components analysis clearly separated Brassica rapa, B. oleracea, B. napus, B. carinata and B. juncea samples into distinct species groups. Several lines were further validated using chromosome counts. Overall, 18% of samples (32/180) were misclassified on the basis of species. Within these 180 samples, 23/76 (30%) supplied on the basis of suspected misclassification were misclassified, and 9/105 (9%) of the samples randomly sourced from the Australian Grains Genebank were misclassified. Surprisingly, several individuals were also found to be the product of interspecific hybridization events. The SNP (single nucleotide polymorphism) array proved effective at confirming species, and provided useful information related to genetic diversity. As similar genomic resources become available for different crops, high-throughput molecular genotyping will offer an efficient and cost-effective method to screen germplasm collections worldwide, facilitating more effective use of these valuable resources by breeders and researchers.
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