Effect of cholangitis after Kasai's portoenterostomy and prognostic fators on the molecular profiles of fecal bacterial flora in an infant with biliary atresia

利用宽范围引物进行16 S rDNA（核糖体RNA基因）PCR和测序的分子方法已被评估其在诊断细菌学中的实用性。首先，使用16 S rRNA基因文库和末端限制性片段长度多态性（T-RFLP）比较来自患有复发性胆管炎的BA个体的粪便内容物中的微生物群。第二，使用16 S rRNA基因文库和T-RFLP比较手术时来自胆总管囊肿（CC）和胆道闭锁（BA）个体的粪便内容物中的微生物群。结果，使用PCR在CC和BA患者中未检测到任何胆汁微生物群（细菌DNA）。细菌DNA显示出显着的差异，粪便微生物群组成的BA婴儿反复胆管炎。利用16 S rRNA基因文库和T-RFLP技术对结肠微生物群进行分子分析可能有助于更早地检测BA胆管炎。细菌DNA显示CC和BA婴儿的粪便微生物群组成存在显着差异。微生物群由双歧杆菌属、乳杆菌目、拟杆菌属、普雷沃氏菌属、梭菌簇IV、Xi和XVIII以及梭菌亚簇XIVa的微生物群落组成。双歧杆菌、类杆菌和梭菌群在CC组中的检出率高于BA组。粪中乳酸杆菌是最主要的菌群。使用16 S rRNA基因文库和T-RFLP对结肠微生物群进行分子分析可能有助于区分CC和BA。

Molecular methods utilizing broad-range primers for 16S rDNA(ribosomal RNA gene) PCR and sequencing have been evaluated for their utility in diagnostic bacteriology. First, microbiota in fecal content from BA individual with recurrent cholangitis was compared using 16S rRNA gene libraries and terminal restriction fragment length polymorphism(T-RFLP). Second, microbiota in fecal content from choledochal cyst(CC) and biliary atresia(BA) individuals at the operation were compared using 16S rRNA gene libraries and T-RFLP As the results, any bile microbiota(bacterial DNA) was not detected using PCR in CC and BA patients. Bacterial DNA showed marked differences in the composition of fecal microbiota in a BA infant with recurrent cholangitis. Molecular analysis of colonic microbiota using 16S rRNA gene libraries and T-RFLP might be useful to detect BA cholangitis more earlier. Bacterial DNA showed marked differences in the composition of fecal microbiota in CC and BA infants. The microbiota consisted of microbial communities of Bifidobacterium, Lactobacillales, Bacteroides, Prevotella, Clostridium clusters IV, XI, and XVIII, and Clostridium subcluster XIVa. The Bifidobacterium, Bacteroides, and Clostridium clusters were detected predominatly in CC than BA group. Lactobacillales was most predominat group in BA feces. Molecular analysis of colonic microbiota using 16S rRNA gene libraries and T-RFLP might be useful to differentiate CC from BA.