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首页> 外文期刊>Journal of Clinical Microbiology >Electron microscopy procedure influences detection of rotaviruses.
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Electron microscopy procedure influences detection of rotaviruses.

机译：电子显微镜检查程序会影响轮状病毒的检测。

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Technical parameters of electron microscope staining procedures (type of stain, pH of stain, and time of staining) influence particle integrity for three groups of rotaviruses. Simian rotavirus SA11 (group A), Chinese adult diarrhea rotavirus and porcine rotavirus-like agent (group B), and porcine pararotavirus (group C) were tested. All rotavirus strains were quite stable in uranyl acetate and phosphotungstic acid at pH 4.5 and relatively stable in ammonium molybdate. However, staining with phosphotungstic acid at higher pH values with increased staining time yielded a reduction in the number of particles and particles that were broken or degraded to single-shelled particles or core particles. The different staining procedures were also tested in immunoelectron microscopy experiments. Antibody molecules bound to rotavirus particles were observed clearly only with phosphotungstic acid staining and not with uranyl acetate. We therefore recommend that uranyl acetate and phosphotungstic acid at pH 4.5 be used for negative staining of rotaviruses; phosphotungstic acid at pH 4.5 is optimal for immunoelectron microscopy. These technical points may be critical for rotavirus detection and are important for studies pertaining to the epidemiology and clinical importance of the non-group A rotaviruses.
机译：电子显微镜染色程序的技术参数（染色类型，染色pH和染色时间）会影响三组轮状病毒的颗粒完整性。测试了猿猴轮状病毒SA11（A组），中国成人腹泻轮状病毒和猪轮状病毒样药（B组）和猪副轮状病毒（C组）。所有轮状病毒株在pH为4.5的乙酸铀酰和磷钨酸中均相当稳定，而在钼酸铵中则相对稳定。但是，在较高的pH值下用磷钨酸进行染色并增加染色时间会导致破碎或降解为单壳粒子或核粒子的粒子数量减少。还在免疫电子显微镜实验中测试了不同的染色程序。仅用磷钨酸染色清楚地观察到与轮状病毒颗粒结合的抗体分子，而没有用乙酸铀酰观察到。因此，我们建议将pH为4.5的乙酸铀酰和磷钨酸用于轮状病毒的阴性染色。 pH 4.5的磷钨酸是免疫电子显微镜的最佳选择。这些技术要点对于轮状病毒的检测可能至关重要，对于与非A型轮状病毒的流行病学和临床重要性有关的研究也很重要。

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