比较Epsilometer试验法和琼脂稀释法检测幽门螺杆菌对甲硝唑的敏感性

doi:10.19723/j.issn.1671-167X.2023.05.024

摘要/Abstract

摘要：

目的: 以琼脂稀释法为金标准, 评价Epsilometer试验(Epsilometer test, E-test)法检测幽门螺杆菌(Helicobacter pylori, H. pylori)对甲硝唑敏感性的一致性。方法: 纳入2018年8月至2020年7月因消化不良症状就诊于北京大学第三医院行胃镜检查的H. pylori感染初治患者, 取胃黏膜组织活检行H. pylori培养, 分别采用E-test法和琼脂稀释法检测H. pylori对甲硝唑的敏感性, 比较两种方法检测结果的一致性和相关性。结果: 成功培养105株H. pylori, 将最小抑菌浓度≥ 8 mg/L定义为耐药。琼脂稀释法检测甲硝唑耐药菌株68株, 耐药率64.8%, E-test法检测耐药菌株66株, 耐药率62.9%, 其中, 琼脂稀释法和E-test法检测均为耐药的菌株66株, 均为敏感的菌株37株, 两种方法的一致率为98.1%。2例菌株被琼脂稀释法评价为耐药, 而E-test法评价为敏感, 非常严重错误率为1.9%。没有菌株被琼脂稀释法评价为敏感, 而E-test法评价为耐药(严重错误率为0%)。以琼脂稀释法为金标准, E-test法检测甲硝唑耐药的灵敏度为97.1%(95%CI: 0.888~0.995), 特异度为100%(95%CI: 0.883~1.000)。Cohen’s kappa系数为0.959 (95%CI: 0.902~1.016, P < 0.001), Spearmans相关性检测r=0.807(P < 0.001)。采用Bland-Altman法进行一致性评价, 结果提示较好, 未出现一致性区间外的测值。E-test法比琼脂稀释法的成本更低, 平均完成1例试验两者的成本分别为269.8元和356.6元。结论: E-test法检测H. pylori对甲硝唑的药敏试验与琼脂稀释法相比具有较强的一致性, E-test法省时、省力、价廉, 可以作为H. pylori药敏试验的优选检测方法。

关键词: Epsilometer试验, 琼脂稀释计数, 幽门螺杆菌, 微生物敏感性试验

Abstract:

Objective: Agar dilution method (ADM) was used as the golden standard to evaluate the consistency of Epsilometer test (E-test) in detecting the sensitivity of Helicobacter pylori (H. pylori) to metronidazole. Methods: From August 2018 to July 2020, patients with H. pylori infection treated for the first time in Peking University Third Hospital for gastroscopy due to dyspepsia were included in this study. Gastric mucosas were taken from the patients with H. pylori infection. H. pylori culture was performed. Both the ADM and E-test were applied to the antibiotic susceptibility of H. pylori to metro-nidazole, and the consistency and correlation between the two methods were validated. Results: In the study, 105 clinical isolates of H. pylori were successfully cultured, and the minimum inhibitory concentration ≥ 8 mg/L was defined as drug resistance. Both ADM and the E-test showed high resistance rates to metronidazole, 64.8% and 62.9%, respectively. Among them, 66 drug-resistant strains were detected by ADM and E-test, and 37 were sensitive strains, so the consistency rate was 98.1%. Two strains were evaluated as drug resistance by ADM, but sensitive by the E-test, with a very major error rate of 1.9%. There was zero strain sensitive according to ADM but assessed as resistant by the E-test, so the major error rate was 0%. Taking ADM as the gold standard, the sensitivity of E-test in the detection of metronidazole susceptibility was 97.1% (95%CI: 0.888-0.995), and the specificity was 100% (95%CI: 0.883-1.000). Cohen's kappa analysis showed substantial agreement, and kappa coefficient was 0.959 (95%CI: 0.902-1.016, P < 0.001). Spearmans correlation analysis confirmed this correlation was significant (r=0.807, P < 0.001). The consistency evaluation of Bland-Altman method indicated that it was good, and there was no measured value outside the consistency interval. In this study, cost analysis, including materials and labor, showed a 32.2% higher cost per analyte for ADM as compared with the E-test (356.6 yuan vs. 269.8 yuan). Conclusion: The susceptibility test of H. pylori to metronidazole by E-test presents better agreement with ADM. Because it is less expensive, less labor intensive, and more rapid, it is an easy and reliable method for H. pylori susceptibility testing.

Key words: Epsilometer test, Agar dilution count, Helicobacter pylori, Microbial sensitivity tests

中图分类号:

R446.5

田雪丽,宋志强,索宝军,周丽雅,李彩玲,张雨欣. 比较Epsilometer试验法和琼脂稀释法检测幽门螺杆菌对甲硝唑的敏感性[J]. 北京大学学报（医学版）, 2023, 55(5): 934-938.

Xue-li TIAN,Zhi-qiang SONG,Bao-jun SUO,Li-ya ZHOU,Cai-ling LI,Yu-xin ZHANG. Comparison of Epsilometer test and agar dilution method in detecting the sensitivity of Helicobacter pylori to metronidazole[J]. Journal of Peking University (Health Sciences), 2023, 55(5): 934-938.

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参考文献 20

1	Sugano K , Tack J , Kuipers EJ , et al. Kyoto global consensus report on Helicobacter pylori gastritis[J]. Gut, 2015, 64 (9): 1353- 1367. doi: 10.1136/gutjnl-2015-309252
2	Malfertheiner P , Megraud F , OMorain CA , et al. Management of Helicobacter pylori infection: the Maastricht V/Florence consensus report[J]. Gut, 2017, 66 (1): 6- 30. doi: 10.1136/gutjnl-2016-312288
3	Hu Y , Zhu Y , Lu NH . Primary antibiotic resistance of Helicobac-ter pylori in China[J]. Dig Dis Sci, 2017, 62 (5): 1146- 1154. doi: 10.1007/s10620-017-4536-8
4	FitzGerald R, Smith SM. An overview of Helicobacter pylori infection [M]// Methods in Molecular Biology. New York: Humana Press, 2021: 1-14.
5	Thung I , Aramin H , Vavinskaya V , et al. Review article: The global emergence of Helicobacter pylori antibiotic resistance[J]. Aliment Pharmacol Ther, 2016, 43 (4): 514- 533. doi: 10.1111/apt.13497
6	Pan J , Shi Z , Lin D , et al. Is tailored therapy based on antibiotic susceptibility effective? A multicenter, open-label, randomized trial[J]. Front Med, 2020, 14 (1): 43- 50. doi: 10.1007/s11684-019-0706-8
7	Alarcon T , Domingo D , Lopez-Brea M . Discrepancies between E-test and agar dilution methods for testing metronidazole susceptibi-lity of Helicobacter pylori[J]. J Clin Microbiol, 1998, 36 (4): 1165- 1166. doi: 10.1128/JCM.36.4.1165-1166.1998
8	Hachem CY , Clarridge JE , Reddy R , et al. Antimicrobial susceptibility testing of Helicobacter pylori, comparison of E-test, broth microdilution, and disk diffusion for ampicillin, clarithromycin, and metronidazole[J]. Diagn Microbiol Infect Dis, 1996, 24 (1): 37- 41. doi: 10.1016/0732-8893(95)00252-9
9	Miftahussurur M , Fauzia KA , Nusi IA , et al. E-test versus agar dilution for antibiotic susceptibility testing of Helicobacter pylori: A comparison study[J]. BMC Res Notes, 2020, 13 (1): 22. doi: 10.1186/s13104-019-4877-9
10	CL SI . Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically[M]. 10th ed Wayne: Clinical and Laboratory Standards Institute, 2015: M07.
11	Alarcón T , Urruzuno P , Martínez MJ , et al. Antimicrobial susceptibility of 6 antimicrobial agents in Helicobacter pylori clinical isolates by using EUCAST breakpoints compared with previously used breakpoints[J]. Enferm Infecc Microbiol Clin, 2017, 35 (5): 278- 282. doi: 10.1016/j.eimc.2016.02.010
12	Hooi JKY , Lai WY , Ng WK , et al. Global prevalence of Helicobacter pylori infection: Systematic review and meta-analysis[J]. Gastroenterology, 2017, 153 (2): 420- 429. doi: 10.1053/j.gastro.2017.04.022
13	刘文忠, 谢勇, 陆红, 等. 第五次全国幽门螺杆菌感染处理共识报告[J]. 中华内科杂志, 2017, 56 (7): 532- 545. doi: 10.3760/cma.j.issn.0578-1426.2017.07.014
14	Mégraud F , Lehn N , Lind T , et al. Antimicrobial susceptibility testing of Helicobacter pylori in a large multicenter trial: The MACH2 study[J]. Antimicrob Agents Chemother, 1999, 43 (11): 2747- 2752. doi: 10.1128/AAC.43.11.2747
15	Glupczynski Y , Broutet N , Cantagrel A , et al. Comparison of the E-test and agar dilution method for antimicrobial suceptibility testing of Helicobacter pylori[J]. Eur J Clin Microbiol Infect Dis, 2002, 21 (7): 549- 552. doi: 10.1007/s10096-002-0757-6
16	Osato MS , Reddy R , Reddy SG , et al. Comparison of the E-test and the NCCLS-approved agar dilution method to detect metro-nidazole and clarithromycin resistant Helicobacter pylori[J]. Int J Antimicrob Agents, 2001, 17 (1): 39- 44. doi: 10.1016/S0924-8579(00)00320-4
17	El-Halfawy OM , Valvano MA . Antimicrobial heteroresistance: An emerging field in need of clarity[J]. Clin Microbiol Rev, 2015, 28 (1): 191- 207. doi: 10.1128/CMR.00058-14
18	Ogata SK , Gales AC , Kawakami E . Antimicrobial susceptibility testing for Helicobacter pylori isolates from Brazilian children and adolescents: Comparing agar dilution, E-test, and disk diffusion[J]. Braz J Microbiol, 2015, 45 (4): 1439- 1448.
19	Best LM , Haldane DJ , Keelan M , et al. Multilaboratory comparison of proficiencies in susceptibility testing of Helicobacter pylori and correlation between agar dilution and E-test methods[J]. Antimicrob Agents Chemother, 2003, 47 (10): 3138- 3144. doi: 10.1128/AAC.47.10.3138-3144.2003
20	Valdivieso-García A , Imgrund R , Deckert A , et al. Cost analysis and antimicrobial susceptibility testing comparing the E-test and the agar dilution method in Campylobacter jejuni and Campylobacter coli[J]. Diagn Microbiol Infect Dis, 2009, 65 (2): 168- 174. doi: 10.1016/j.diagmicrobio.2009.07.008

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	E-test			ADM
	Unit-price/yuan	n	Total/yuan	Unit-price/yuan	n	Total/yuan
E-test strip	35.0	1	35.0		-
Agar dilution plate (11 gradients)		-		8.8	11	96.8
Plate (containing antibiotics)	12.0	2	24.0	12.0	2	24.0
Plate (without antibiotics)	8.8	4	35.2	8.8	4	35.2
General culture plate	25.0	2	50.0	25.0	2	50.0
Nitrogen, water and electricity	60.0	1	60.0	60.0	1	60.0
Manual labor	50.0	1	50.0	75.0	1	75.0
Others			15.6			15.6
Total			269.8			356.6

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