wzi测序在粪便携带肺炎克雷伯菌的血清型检测中的应用
| 作者: |
1范沁榕,
1沈兰凤,
1胡仁静
1 南京医科大学附属无锡市第二人民医院检验科，江苏 无锡 214002 |
| 通讯: |
胡仁静
Email: weiweihuhu112@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2020.11.007 |
| 基金: | 无锡市科技支撑项目(CSE31N1602)；无锡市卫生健康科研项目计划(Q201901)。 |
摘要
目的：了解糖尿病患者粪便中肺炎克雷伯菌的荚膜血清型分布，建立wzi基因测序检测肺炎克雷伯菌荚膜血清型的体系。方法：收集无锡市第二人民医院2018年1月至2018年12月共370份粪便标本，其中糖尿病组120份，腹泻组135份，健康体检组115份。所有标本进行培养进一步分离肺炎克雷伯菌。采用普通PCR筛查肺炎克雷伯菌常见的10种高毒力荚膜血清型(K1，K2，K5，K16，K20，K54，K57，KN1，KN2，KN3)和4种常见的非高毒力荚膜血清型(K3，K15，K47，K64)；同时全部菌株进行wzi基因测序检测荚膜血清型。结果：粪便标本中肺炎克雷伯菌的总分离率为25.41%(94/370)；糖尿病患者分离率为35.00%(42/120)，显著高于腹泻患者的19.26%(26/135)和对照组健康体检者的22.61%(26/115；χ2=8.995，P=0.01)。wzi基因测序可检出90.43%(85/94)菌株的荚膜血清型共30种，普通PCR选择的14种荚膜血清型仅检测出其中的11种；糖尿病患者组高毒力肺炎克雷伯菌的检出率为61.90%(26/42)高于腹泻患者组26.92%(7/26)和健康体检组46.15%(12/26；χ2=7.917，P=0.019)。糖尿病患者组中检出的高毒力血清型以K1，K2型为主，占56.69%(15/26)。结论：wzi基因测序可高效检测肺炎克雷伯菌的荚膜血清型；糖尿病患者粪便中高毒力肺炎克雷伯菌分离率显著高于腹泻患者和对照组健康体检者，需加重视对糖尿病患者粪便中定植的肺炎克雷伯菌的筛查。
关键词：
肺炎克雷伯菌；wzi基因测序；荚膜血清型；糖尿病
Application of wzi gene sequencing in the detection of serotypes for Klebsiella pneumoniae in feces
CorrespondingAuthor: HU Renjing Email: weiweihuhu112@163.com
DOI: 10.3978/j.issn.2095-6959.2020.11.007
Foundation: This work was supported by the Wuxi Science and Technology Development Fund Project (CSE31N1602); Wuxi Health and Scientific Research Project Plan (Q201901), China.
Abstract
Objective: To understand the distribution of capsular serotypes of Klebsiella pneumoniae in feces of diabetic patients, and to establish a system for detecting Klebsiella pneumoniae capsule serotypes by wzi gene sequencing. Methods: A total of 370 fecal samples were collected from January 2018 to December 2018 in the Second People’s Hospital of Wuxi, including 120 in the diabetes group, 135 in the diarrhea group and 115 in the health and physical examination group. All specimens were cultured for further isolation of Klebsiella pneumoniae. Ten common high virulent capsular serotypes (K1, K2, K5, K16, K20, K54, K57, KN1, KN2, KN3) and four non-high virulent capsular serotypes (K3, K15, K47, K64) of Klebsiella pneumoniae were detected by common Polymerase Chain Reaction (PCR). At the same time, wzi gene sequencing of capsular serotypes of all strains was carried out simultaneously. Results: The total isolation rate of Klebsiella pneumoniae in fecal specimens was 25.41% (94/370). The separation rate of diabetic patients was 35.00% (42/120), which was higher than that of diarrhea patients 19.26% (26/135) and control group 22.61% (26/115) (χ2=8.995, P=0.01). Wzi gene sequencing detected a total of 30 capsular serotypes of 90.43% (85/94) strain, and only 11 of the 14 capsular serotypes selected by common PCR were detected. The detection rate of hypervirulent Klebsiella pneumoniae in the diabetic group was 61.90% (26/42), which was higher than that in the diarrhea group 26.92%(7/26) and the healthy physical examination group 46.15% (12/26; χ2=7.917, P=0.019). The high virulence serotypes detected in the diabetic group were mainly K1 and K2, accounting for 56.69% (15/26) of hypervirulent Klebsiella pneumoniae. Conclusion: wzi gene sequencing can efficiently detect the capsular serotype of Klebsiella pneumoniae; the isolation rate of Klebsiella pneumoniae in the feces of diabetic patients with high virulence was significantly higher than that of diarrhea patients and the control group. Therefore, more attention should be paid to the screening of Klebsiella pneumoniae colonized in the feces of diabetic patients.
Keywords：
Klebsiella pneumoniae; wzi gene sequencing; capsular serotype; diabetes mellitus