González-Escalona Narjol, Gavilan Ronnie G, Brown Eric W, Martinez-Urtaza Jaime
Center for Food and Applied Nutrition, Food and Drug Administration, College Park, MD, United States of America.
Centro Nacional de Salud Publica, Instituto Nacional de Salud, Lima, Peru.
PLoS One. 2015 Feb 13;10(2):e0117485. doi: 10.1371/journal.pone.0117485. eCollection 2015.
Vibrio parahaemolyticus is an important human pathogen whose transmission is associated with the consumption of contaminated seafood. Consistent multilocus sequence typing for V. parahaemolyticus has shown difficulties in the amplification of the recA gene by PCR associated with a lack of amplification or a larger PCR product than expected. In one strain (090-96, Peru, 1996), the produced PCR product was determined to be composed of two recA fragments derived from different Vibrio species. To better understand this phenomenon, we sequenced the whole genome of this strain. The hybrid recA gene was found to be the result of a fragmentation of the original lineage-specific recA gene resulting from a DNA insertion of approximately 30 kb in length. This insert had a G+C content of 38.8%, lower than that of the average G+C content of V. parahaemolyticus (45.2%), and contained 19 ORFs, including a complete recA gene. This new acquired recA gene deviated 24% in sequence from the original recA and was distantly related to recA genes from bacteria of the Vibrionaceae family. The reconstruction of the original recA gene (recA3) identified the precursor as belonging to ST189, a sequence type reported previously only in Asian countries. The identification of this singular genetic feature in strains from Asia reveals new evidence for genetic connectivity between V. parahaemolyticus populations at both sides of the Pacific Ocean that, in addition to the previously described pandemic clone, supports the existence of a recurrent transoceanic spreading of pathogenic V. parahaemolyticus with the corresponding potential risk of pandemic expansion.
副溶血性弧菌是一种重要的人类病原体,其传播与食用受污染的海鲜有关。对副溶血性弧菌进行一致的多位点序列分型时,通过PCR扩增recA基因存在困难,表现为缺乏扩增产物或出现比预期更大的PCR产物。在一株菌株(090 - 96,秘鲁,1996年)中,所产生的PCR产物被确定为由来自不同弧菌物种的两个recA片段组成。为了更好地理解这一现象,我们对该菌株的全基因组进行了测序。发现杂交recA基因是原始谱系特异性recA基因片段化的结果,该片段化是由一个长度约为30 kb的DNA插入导致的。这个插入片段的G + C含量为38.8%,低于副溶血性弧菌的平均G + C含量(45.2%),并且包含19个开放阅读框,其中包括一个完整的recA基因。这个新获得的recA基因在序列上与原始recA基因有24%的偏差,并且与弧菌科细菌的recA基因亲缘关系较远。对原始recA基因(recA3)的重建确定其前体属于ST189,这是一种此前仅在亚洲国家报道过的序列类型。在亚洲菌株中鉴定出这种独特的遗传特征,揭示了太平洋两岸副溶血性弧菌种群之间遗传联系的新证据,这除了先前描述的大流行克隆外,还支持致病性副溶血性弧菌存在反复的跨洋传播以及相应的大流行扩展潜在风险。