| Objective:To study the major virulence genes and the drug resistance of Helicobacter pylori(Hp)in gastric mucosa and saliva of patients with gastrointestinal symptoms by the PCR-reverse dot bot(PCR-RDB)in the First Hospital of Jilin University,and compare the similarities and differences of results between gastric mucosa and saliva.Methods:Rapid Urease Test(RUT)was performed on gastric mucosa of patients undergoing gastroscopy at the Endoscopy Center of the First Hospital of Jilin University,collecting gastric mucosa specimens from 169 patients and saliva specimens from 100 patients with RUT positive.PCR-RDB was used to detect Hp DNA,major virulence genes and the mutations of drug resistance genes.Result:1.The infection and drug resistance of Hp in the First Hospital of Jilin University:(1)The positive rate of Hp DNA in 169 gastric mucosa specimens was 69.2%(117/169).(2)Three types of vac A genotypes were detected: s1m1(37.6%),s1m2(59.8%),s1m1 and s1m2 coexist(2.6%).There was no significant differences between vac A genotypes and disease types(P > 0.05).(3)Among 117 gastric mucosa specimens with Hp DNA positive,the drug resistance rate of clarithromycin was 71.8%(84/117);levofloxacin was 41.9%(49/117);metronidazole was 35%(41/117);Tetracycline was 17.1%(20/117);Amoxicillin was 4.3%(5/117).(4)The drug resistance patterns of Hp: single drug resistance rate was 26.5%,and clarithromycin was the highest(14.5%);double drug resistance rate was 35.9%,and clarithromycin + levofloxacin group was the highest(17.1%);multi-drug resistance rate was 23.9%,with the highest in the clarithromycin + levofloxacin + metronidazole group(11.1%).(5)The drug resistance rate of metronidazole in male patients was higher than that in female patients,the difference was statistically significant(P = 0.006).The drug resistance rates of clarithromycin,metronidazole,levofloxacin and tetracycline in different age groups were different,there were statistically significances(P <0.05).The drug resistance rates of clarithromycin,levofloxacin and tetracycline have the characteristics of high age group> low age group.The resistance rates of clarithromycin,metronidazole,levofloxacin and tetracycline in different diseases were different,and the resistance rate of non-atrophic gastritis was the highest,there were statistically differences(P <0.05).(6)The mutation sites of amoxicillin resistance in Hp were mainly T556 S of pbp1 gene(80%),A2143 G of 23 S r RNA gene in clarithromycin(97.6%),G616 A of rdx A gene in metronidazole(100%),D91G(38.8%)and N87K(30.6%)of gyr A gene in levofloxacin,and AG926-927 GT of 16 S r RNA gene in tetracycline(65%).2.Comparison of Hp infection and drug resistance in gastric mucosa and saliva specimens of 100 patients:(1)The positive rate of Hp DNA in gastric mucosa was 77.0%(77/100),and saliva was 39.0%(39/100),the difference was statistically significant(P <0.001).(2)The positive rate of vac A genotypes in gastric mucosa was 35.1%(27/77)in s1m1,62.3%(48/77)in s1m2,s1m1 and s1m2 coexist was 2.6%(2/77).S1m1 in saliva specimen was 38.5%(15/39),s1m2 was 61.5%(24/39).There were no statistically significant differences(P> 0.05).(3)The drug resistance rates of gastric mucosa and saliva specimens were 90.9%(70/77)and 92.3%(36/39),respectively,and no statistical differences in drug resistance rates(P> 0.05).There were also no significances in the single,double and multi-drug resistance rates of Hp between gastric mucosa and saliva(all P> 0.05).(4)The major mutation sites of Hp drug resistance genes in gastric mucosa and saliva samples were: T556 S,A2143G,G616 A,D91G,N87 K,and AG926-927 GT.The mutation rates of drug resistance genes in gastric mucosa and saliva were not statistically different(P> 0.05).(5)38 gastric mucosa and saliva samples were simultaneously positive of Hp DNA,and vac A genotypes were completely consistent.The coincidence rate of mutation sites was 68.4%(26/38).One patient’s gastric mucosa specimen was negative for Hp DNA,while the saliva specimen was positive.Conclusion:1.In the First Hospital of Jilin University,vac A genotypes of Hp in gastric mucosa were mainly s1m1 and s1m2,which had strong virulence,there were no correlation between genotypes and disease types.There were both s1m1 and s1m2 in some patients,suggesting that there was mixed infection of Hp.The drug resistance rates of Hp to clarithromycin,metronidazole and levofloxacin were high.The drug resistance rate of metronidazole in male patients was higher.The resistance rates of clarithromycin,levofloxacin and tetracycline were higher in the older age group.Patients with non-atrophic gastritis have a higher drug resistance rate except amoxicillin.Hp resistance patterns were diverse,double and multi-drug resistance rates were high,and drug resistance was serious.The major mutation sites of drug resistance genes of Hp in the First Hospital of Jilin University were: T556 S,A2143G,G616 A,D91G,N87 K,AG926-927 GT.2.There were no differences in drug resistance rates of Hp and mutation rates of drug resistance genes in gastric mucosa and saliva.The mutation sites of drug resistance genes in gastric mucosa and saliva were not completely consistent.The detection rate of Hp in gastric mucosa was significantly higher than that in saliva.One patient’s gastric mucosa specimen was negative for Hp DNA,while the saliva specimen was positive.Combined detection of Hp in gastric mucosa and saliva can improve the detection rate of Hp in patients. |
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