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一种新型 mba 实时 PCR 方法用于蒙古母亲及其后代中 Ureaplasma parvum 的基因分型验证。

A novel mba-based Real time PCR approach for genotyping of Ureaplasma parvum validated in a cohort of Mongolian mothers and offspring.

机构信息

Department of Microbiology and Immunology, School of Biomedicine, Mongolian National University of Medical Sciences, Zorig Street, Ulaanbaatar, 14210, Mongolia.

Institute of Medical Microbiology and Hospital Hygiene, University Hospital, Heinrich-Heine-University Düsseldorf, University Street, Düsseldorf, 40225, Germany.

出版信息

Int J Med Microbiol. 2018 Oct;308(7):865-871. doi: 10.1016/j.ijmm.2018.08.001. Epub 2018 Aug 6.

Abstract

The role of Ureaplasma parvum in abnormal outcomes of human pregnancy has been discussed controversially in the past. Of the 14 known ureaplasma serovars, the Ureaplasma parvum serovars 1, 3, 6 and 14, have been found to derive from smaller genomes. Serovars 3 and 6 have been described more often to cause complications in pregnancy. To elucidate the serovar distribution in U. parvum positive specimens of 200 Mongolian mothers and their offspring, a new set of mba-targeting PCRs was developed enabling a fast and reliable serovar differentiation by melting peak analysis in a Real time PCR approach or by conventional agarose gel electrophoresis. 92% maternal and 55% neonatal samples were retrospectively genotyped and a dominance of serovars 3 and 6 was detected while serovar 14 was almost absent. Transmission from mothers to newborns was detected in 83% of U. parvum positive neonates exhibiting serovar patterns identical to their mothers. No statistically significant correlation between a distinct serovar and pregnancy outcome could be detected. However, neonatal colonization with serovar 1 declined with progressing pregnancy suggesting that a higher ureaplasma load shortened pregnancy and thereby had a potential negative effect on offspring health. Our novel mba-based Real time PCR approach, which can also be used in conventional PCR and gel electrophoretic analysis, provides the proof of principle that the four U. parvum serovars 1, 3, 6 and 14 can be differentially detected and quantified. A larger scale study outside the scope of this work should be conducted to clarify the impact of serovar 1 on pregnancy outcome.

摘要

过去,关于微小脲原体(Ureaplasma parvum)在人类妊娠异常结局中的作用存在争议。在已知的 14 种脲原体血清型中,Ureaplasma parvum 血清型 1、3、6 和 14 的基因组较小。血清型 3 和 6 更常被描述为导致妊娠并发症。为了阐明 200 位蒙古母亲及其后代的 U. parvum 阳性标本中的血清型分布,开发了一套新的 mba 靶向 PCR,通过实时 PCR 方法中的熔解峰分析或传统琼脂糖凝胶电泳,可快速可靠地区分血清型。92%的母本和 55%的新生儿样本进行了回顾性基因分型,发现血清型 3 和 6 占优势,而血清型 14 几乎不存在。在 83%的 U. parvum 阳性新生儿中检测到从母亲到新生儿的传播,其血清型模式与母亲相同。未检测到特定血清型与妊娠结局之间存在统计学显著相关性。然而,随着妊娠的进展,新生儿血清型 1 的定植减少,这表明脲原体负荷较高会缩短妊娠时间,从而对后代健康产生潜在的负面影响。我们基于 mba 的新型实时 PCR 方法也可用于常规 PCR 和凝胶电泳分析,为可以区分和定量检测四种 U. parvum 血清型 1、3、6 和 14 提供了原理证明。应进行更大规模的、本研究范围之外的研究,以阐明血清型 1 对妊娠结局的影响。

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