School of Molecular & Microbial Biosciences, University of Sydney, Sydney, NSW, Australia.
J Virol Methods. 2010 May;165(2):178-85. doi: 10.1016/j.jviromet.2010.01.015. Epub 2010 Feb 1.
The use of high resolution mass spectrometry to detect signature peptides within proteolytic digests of the isolated matrix M1 protein, and whole virus digests, for both human and animal strains of influenza is shown to be able to rapidly and reliably type the virus. Conserved sequences for predicted tryptic peptides were identified through alignments of matrix M1 protein sequences across all human, avian and swine strains of the influenza virus. Peptides with unique masses, when compared with those from the in silico digestion of all influenza antigens and those proteins known to contaminate egg grown strains, were identified using the purpose built FluGest algorithm. Their frequency of occurrence within the matrix M1 protein across all type A and type B strains was established with the FluAlign algorithm. The subsequent detection of the signature peptides of matrix M1 protein within proteolytic digests of type A and type B human and avian strains has been demonstrated.
使用高分辨率质谱法检测分离的基质 M1 蛋白以及整个病毒消化物中的肽段特征，可以快速、可靠地对病毒进行分型。通过对所有人类、禽和猪流感病毒株的基质 M1 蛋白序列进行比对，确定了预测胰蛋白酶肽段的保守序列。使用专门设计的 FluGest 算法，通过与所有流感抗原的计算机消化肽段以及已知污染鸡蛋生长株的蛋白质进行比较，确定了具有独特质量的肽段。使用 FluAlign 算法确定了它们在所有 A 型和 B 型株的基质 M1 蛋白中的出现频率。随后，已经证明可以在 A 型和 B 型人类和禽流感株的蛋白水解消化物中检测到基质 M1 蛋白的特征肽段。
