Real-time PCR for sexing Schistosoma mansoni cercariae.
Real-time PCR for sexing Schistosoma mansoni cercariae.
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DOI：
10.1016/j.molbiopara.2016.03.010
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发表时间：
2016-01
影响因子：
1.5
通讯作者：
Anderson TJ
中科院分区：
文献类型：
--
作者：
Chevalier FD;Le Clec'h W;Alves de Mattos AC;LoVerde PT;Anderson TJ
尾蚴幼虫的性别只能用分子方法来确定。已经开发了扩增雌性（ZW）寄生虫W染色体上重复标记的终点PCR方法，但有时结果不明确或不正确。为了更有效地区分性别，并确定为什么终点PCR可能是不正确的，我们使用实时PCR定量W 6重复序列和特定的Z染色体基因。W 6和Z染色体标记的拷贝数之间的比率明确识别性别：女性具有比男性（0-21）更高的比率（421-4371）。然而，一些男性在其基因组中具有低数量的W 6元件，并且与给出明确终点结果的男性相比，qPCR显示出给出模糊终点PCR结果的男性基因型的W 6/Z标记比率显著更高。定量PCR性别鉴定方法的发展将是特别有用的，可靠的尾蚴性别鉴定是至关重要的，例如当分期遗传杂交。
The gender of cercarial larvae can only be determined using molecular methods. End point PCR methods that amplify repetitive markers on the W chromosome of the female (ZW) parasites have been developed, but sometimes results are ambiguous or incorrect. To more effectively distinguish sexes, and to determine why end point PCR can be incorrect, we quantified the W6 repeat sequence and a specific Z chromosome gene using real-time PCR. The ratio between copy number of W6 and a Z chromosome marker unambiguously identifies gender: females have higher ratios (421-4371) than males (0–21). However, some males have low numbers of W6 elements in their genome, and qPCR demonstrated significantly higher W6/Z marker ratios for male genotypes giving ambiguous end point PCR results compared with males giving clear end point results. The quantitative PCR sexing method developed will be particularly useful where reliable sexing of cercariae is critical, for example when staging genetic crosses.