Feberwee Anneke, Dijkstra Jeroen R, von Banniseht-Wysmuller Thea E, Gielkens Arno L J, Wagenaar Jaap A
Poultry Health Centre, Animal Health Service, P.O. Box 9, 7400 AA Deventer, the Netherlands.
Vet Microbiol. 2005 Nov 30;111(1-2):125-31. doi: 10.1016/j.vetmic.2005.09.003. Epub 2005 Oct 19.
Mycoplasma gallisepticum (MG) and M. synoviae (MS) are the cause of considerable economic losses in the poultry industry. Molecular differentiation of avian Mycoplasma strains may be helpful in tracing infections and in the evaluation of implemented intervention strategies. Amplified Fragment Length Polymorphism (AFLP) has shown to be a powerful typing technique but the application for poultry Mycoplasma strains is very limited. The aim of this study was to evaluate the reproducibility and discriminatory power of AFLP HindIII/HhaI and AFLP BglII/Mfel for the inter- and intraspecies differentiation of avian mycoplasmas and to compare these test characteristics with digitalized Random Amplified Polymorphic DNA (RAPD) analysis. The reproducibility of RAPD, AFLP HindIII/HhaI and AFLP BglII/Mfel was 50-100, 97-98 and 86-99%, respectively. RAPD and both AFLP enzyme combinations were able to differentiate between five avian Mycoplasma species. For AFLP, five MG and four MS clusters could be identified. The phylogenetic tree for both enzyme combinations was comparable. For RAPD, four MG clusters could be identified. For MS, however, due to the poor reproducibility of the RAPD technique, no clear genogroups could be identified. On basis of the results of this study it can be concluded that AFLP is a powerful technique for the genotyping of avian mycoplasmas and that, although AFLP HindIII/HhaI generated patterns with less fragments, the final results showed homologous results.
鸡毒支原体（MG）和滑液支原体（MS）是家禽业经济损失的重要原因。禽支原体菌株的分子鉴别有助于追踪感染情况以及评估实施的干预策略。扩增片段长度多态性（AFLP）已被证明是一种强大的分型技术，但在家禽支原体菌株中的应用非常有限。本研究的目的是评估AFLP HindIII/HhaI和AFLP BglII/Mfel用于禽支原体种间和种内鉴别的可重复性和鉴别能力，并将这些测试特征与数字化随机扩增多态性DNA（RAPD）分析进行比较。RAPD、AFLP HindIII/HhaI和AFLP BglII/Mfel的可重复性分别为50 - 100%、97 - 98%和86 - 99%。RAPD和两种AFLP酶组合都能够区分五种禽支原体。对于AFLP，可以识别出五个MG簇和四个MS簇。两种酶组合的系统发育树具有可比性。对于RAPD，可以识别出四个MG簇。然而，对于MS，由于RAPD技术的可重复性较差，无法识别出清晰的基因群。根据本研究结果可以得出结论，AFLP是一种用于禽支原体基因分型的强大技术，并且尽管AFLP HindIII/HhaI产生的条带较少，但最终结果显示出同源结果。
