Méndez-Tenorio Alfonso, Flores-Cortés Perla, Guerra-Trejo Armando, Jaimes-Díaz Hueman, Reyes-Rosales Emma, Maldonado-Rodríguez Arcadio, Espinosa-Lara Mercedes, Maldonado-Rodríguez Rogelio, Kenneth Loren Beattie
Laboratorio de Tecnología del DNA, Escuela Nacional de Ciencias Biológicas. IPN. Prol. de Carpio y Plan de Ayala s/n, Col. Santo Tomás, 11340, Mexico DF.
Rev Latinoam Microbiol. 2006 Apr-Jun;48(2):56-65.
The identification of microorganisms by whole genome DNA fingerprinting was tested "in silico". 94 HPV genome sequences were submitted to virtual hybridization analysis on a DNA chip with 342 probes. This Universal Fingerprinting Chip (UFC) constitutes a representative set of probes of all the possible 8-mer sequences having at least two internal and non contiguous sequence differences between all them. A virtual hybridization analysis was performed in order to find the fingerprinting pattern that represents the signals produced for the hybridization of the probes allowing at most a single mismatch. All the fingerprints for each virus were compared against each other in order to obtain all the pairwise distances measures. A match-extension strategy was applied to identify only the shared signals corresponding to the hybridization of the probes with homologous sequences between two HPV genomes. A phylogenetic tree was constructed from the fingerprint distances using the Neighbor-Joining algorithm implemented in the program Phylip 3.61. This tree was compared with that produced from the alignment of whole genome HPV sequences calculated with the program Clustal_X 1.83. The similarities between both trees are suggesting that the UFC-8 is able to discriminate accurately between viral genomes. A fingerprint comparative analysis suggests that the UFC-8 can differentiate between HPV types and sub-types.
通过全基因组DNA指纹识别微生物的方法在计算机上进行了测试。94个HPV基因组序列被提交至含有342个探针的DNA芯片上进行虚拟杂交分析。这种通用指纹芯片(UFC)由所有可能的8聚体序列的代表性探针集组成,这些序列之间至少有两个内部且不连续的序列差异。进行虚拟杂交分析以找到代表探针杂交产生的信号的指纹图谱,该杂交允许最多一个错配。将每种病毒的所有指纹相互比较,以获得所有成对距离测量值。应用匹配扩展策略仅识别与两个HPV基因组之间同源序列的探针杂交对应的共享信号。使用Phylip 3.61程序中实现的邻接法从指纹距离构建系统发育树。将该树与用Clustal_X 1.83程序计算的全基因组HPV序列比对产生的树进行比较。两棵树之间的相似性表明UFC-8能够准确区分病毒基因组。指纹比较分析表明UFC-8可以区分HPV类型和亚型。