Quantitative detection of Helicobacter pylori gene expression in vivo and relationship to gastric pathology.
Quantitative detection of Helicobacter pylori gene expression in vivo and relationship to gastric pathology.
复制标题
体内幽门螺杆菌基因表达的定量检测及其与胃病理的关系。
DOI：
10.1128/iai.68.10.5488-5495.2000
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发表时间：
2000
影响因子：
3.1
通讯作者：
Miller,GG
中科院分区：
文献类型：
--
作者：
PeekJr,RM;vanDoorn,LJ;Donahue,JP;Tham,KT;Figueiredo,C;Blaser,MJ;Miller,GG
幽门螺杆菌的iceA基因座包括iceA 1和diceA 2两个互斥基因家族之一。与iceA 1菌株定植与急性粘膜炎症增强，并坚持胃上皮细胞在体外诱导表达iceA 1，但ceaseA 2 mRNA，然而，这两个转录本可以在体内检测。本研究的目的是确定体内不同水平的iceA转录是否有助于疾病的发病机制。41 H的RNA幽门阳性胃活检标本逆转录为cDNA。用生物素标记的iceA 1、iceA 2和16 S rRNA引物进行定量PCR，用荧光素标记的探针进行杂交检测生物素标记产物与链霉亲和素包被平板的结合，PCR和序列分析确定iceA基因型。所有41个样品均含有可检测的H。pylori 16 S rRNA在A1（n= 10）和A2（n= 31）定植患者中的水平相似（P= 0.34）。分别有4名（40%）和19名（61%）人的活检标本中存在iceA 1或iceA 2菌株，可检测到iceARNA。iniceA 1 RNA阳性患者的急性炎症评分显著高于iniceA 1 RNA阴性、iceA 2 RNA阳性或iceA 2 RNA阴性受试者（均P≤ 0.05）。在A2 RNA阳性组中，H.具有单个35-氨基酸盒的pylori菌株与显著更高的mucosaliceA 2转录水平相关（与具有两个盒的菌株相比P= 0.014）。这些结果表明，H. pylori iceA 1和iceA 2基因的转录水平与16 SrRNA的转录水平无关，iceA 2基因的特殊结构与转录水平的增强有关。A1转录水平与嗜中性粒细胞浸润强度显著相关的发现表明，在H. pylori iceA 1菌株反映了体内iceA 1转录水平。
TheiceAlocus ofHelicobacter pyloriincludes one of two mutually exclusive gene families,iceA1andiceA2. Colonization withiceA1strains is associated with enhanced acute mucosal inflammation, and adherence to gastric epithelial cells in vitro induces expression oficeA1but noticeA2mRNA; however, both transcripts can be detected in vivo. The aim of this study was to determine whether differing levels oficeAtranscription in vivo may contribute to disease pathogenesis. RNA from 41H. pylori-positive gastric biopsy specimens was reverse transcribed to cDNA. Quantitative PCR was performed using biotinylated iceA1,iceA2, and 16S rRNA primers, and binding of biotinylated products to streptavidin-coated plates was detected by hybridization with a fluorescein-labeled probe.iceAgenotypes were determined by PCR and sequence analysis. All 41 samples contained detectableH. pylori16S rRNA, with similar levels iniceA1- (n= 10) andiceA2(n= 31)-colonized patients (P= 0.34). Biopsy specimens from four (40%) and 19 (61%) persons colonized withiceA1oriceA2strains, respectively, had detectableiceARNA. Acute inflammatory scores were significantly higher iniceA1RNA-positive patients than iniceA1RNA-negative,iceA2RNA-positive, oriceA2RNA-negative subjects (P≤ 0.05 for each). Within theiceA2RNA-positive group,H. pyloristrains with a single 35-amino-acid cassette were associated with significantly higher mucosaliceA2transcript levels (P= 0.014 versus strains with two cassettes). These results indicate that the levels of transcription ofH. pylori iceA1andiceA2and of 16S rRNA are independent and that particulariceA2gene structures are associated with enhanced transcription. The finding thaticeA1transcription levels are significantly associated with the intensity of neutrophilic infiltration suggests that heterogeneity in inflammatory scores among persons colonized withH. pylori iceA1strains reflects levels oficeA1transcription in vivo.