Hanelt B, Steinauer M L, Mwangi I N, Maina G M, Agola L E, Mkoji G M, Loker E S
Department of Biology, University of New Mexico, Albuquerque, NM 87131, USA.
Trop Med Int Health. 2009 Mar;14(3):322-31. doi: 10.1111/j.1365-3156.2009.02226.x. Epub 2009 Jan 28.
To develop and assess a microsatellite technique to characterize populations of Schistosoma mansoni from humans.
For each of five patients, we calculated the allele count and frequency at 11 loci for several pools of miracidia (50 and 100), and compared these to population values, determined by amplifying microsatellites from 186 to 200 individual miracidia per patient.
We were able to detect up to 94.5% of alleles in pools. Allele count and frequency strongly and significantly correlated between singles and pools; marginally significant differences (P < 0.05) were detected for one patient (pools of 50) for allele frequencies and for two patients (pools of 100) for allele counts. Kato-Katz egg counts and number of alleles per pool did not co-vary, indicating that further direct comparisons of the results from these two techniques are needed.
Allele counts and frequency profiles from pooling provide important information about infection intensity and complexity, beyond that obtained from traditional methods. Although we are not advocating use of pooling to replace individual genotyping studies, it can potentially be useful in certain applications as a rapid and cost effective screening method for studies of S. mansoni population genetics, or as a more informative way to quantify and characterize human worm populations.
开发并评估一种微卫星技术,用于对来自人类的曼氏血吸虫种群进行特征分析。
对于五名患者中的每一位,我们计算了几个毛蚴池(50个和100个)在11个位点的等位基因计数和频率,并将这些结果与通过对每位患者186至200个单个毛蚴的微卫星进行扩增所确定的群体值进行比较。
我们能够在池样本中检测到高达94.5%的等位基因。单个样本和池样本之间的等位基因计数和频率呈强且显著的相关性;对于一名患者(50个样本的池)的等位基因频率以及两名患者(100个样本的池)的等位基因计数,检测到了边缘显著差异(P < 0.05)。加藤-凯茨虫卵计数与每个池中的等位基因数量没有共同变化,这表明需要对这两种技术的结果进行进一步的直接比较。
来自池样本的等位基因计数和频率分布提供了有关感染强度和复杂性的重要信息,这超出了传统方法所获得的信息。虽然我们并不主张使用池样本法来取代个体基因分型研究,但它在某些应用中可能是有用的,可作为一种快速且经济高效的筛选方法,用于曼氏血吸虫群体遗传学研究,或者作为一种更具信息量的方式来量化和表征人类蠕虫群体。