Peña J A, Fox J G, Ferraro M J, Versalovic J
Department of Medical Laboratory Science, Northeastern University, Boston, MA, USA.
Arch Pathol Lab Med. 2001 Apr;125(4):493-7. doi: 10.5858/2001-125-0493-MRTOHP.
To evaluate simultaneous diagnosis of infection and molecular resistance testing of Helicobacter pylori.
Gastric biopsies were obtained from 26 rapid urease-positive and 51 rapid urease-negative test kits used to diagnose H pylori infection. Following glass bead-assisted DNA isolation, amplification of H pylori 16S ribosomal DNA (rDNA), glmM, and 23S rDNA target genes was performed.
Helicobacter pylori DNA was successfully amplified from 100% (26/26) of urease-positive and 3.9% (2/51) of urease-negative gastric biopsies. Subsequent restriction enzyme-mediated digestion of 23S rDNA amplification products revealed that 17% (4/24) of urease-positive and H pylori DNA-positive biopsy specimens contained point mutations (A2142G or A2143G) associated with clarithromycin resistance. Helicobacter pylori DNA from gastric biopsies was successfully amplified 8 weeks following rapid urease testing.
Helicobacter pylori genotyping may be used to detect macrolide-resistant H pylori in individuals prior to initiation of therapy or in patients refractory to anti-H pylori therapy. Two urease-negative specimens yielded Helicobacter DNA distinct from that of H pylori and indicated the need for further investigations of Helicobacter species present in the human stomach.
评估幽门螺杆菌感染的同步诊断及分子耐药性检测。
从用于诊断幽门螺杆菌感染的26个快速尿素酶阳性和51个快速尿素酶阴性检测试剂盒中获取胃活检组织。在玻璃珠辅助DNA分离后，对幽门螺杆菌16S核糖体DNA（rDNA）、glmM和23S rDNA靶基因进行扩增。
幽门螺杆菌DNA在100%（26/26）的尿素酶阳性和3.9%（2/51）的尿素酶阴性胃活检组织中成功扩增。随后对23S rDNA扩增产物进行限制性内切酶介导的消化显示，17%（4/24）的尿素酶阳性且幽门螺杆菌DNA阳性的活检标本含有与克拉霉素耐药相关的点突变（A2142G或A2143G）。快速尿素酶检测8周后，胃活检组织中的幽门螺杆菌DNA成功扩增。
幽门螺杆菌基因分型可用于在开始治疗前或对抗幽门螺杆菌治疗难治的患者中检测耐大环内酯类幽门螺杆菌。两个尿素酶阴性标本产生的幽门螺杆菌DNA与幽门螺杆菌不同，表明需要对人胃中存在的幽门螺杆菌种类进行进一步研究。
