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冷冻保存条件下阿莫西林耐药性的遗传和转录组变异

Genetic and Transcriptomic Variations for Amoxicillin Resistance in under Cryopreservation.

作者信息

Han Xiurui, Zhang Yiyao, He Lihua, Fan Ruyue, Sun Lu, Fan Dongjie, Gong Yanan, Chen Xiaoli, You Yuanhai, Zhao Fei, Zhang Maojun, Zhang Jianzhong

机构信息

State Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.

出版信息

Pathogens. 2021 May 30;10(6):676. doi: 10.3390/pathogens10060676.

DOI:10.3390/pathogens10060676
PMID:34070823
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8229390/
Abstract

Some amoxicillin-resistant strains of show a sharp decrease in amoxicillin resistance after freezing. In China, most clinical gastric mucosal specimens are frozen and transported for isolation and drug susceptibility testing for , which may lead to an underestimation of the amoxicillin resistance. The objective of this study is to investigated reasons for the decreased amoxicillin resistance after cryopreservation. A high-level amoxicillin-resistant clone (NX24r) was obtained through amoxicillin pressure screening. After cryopreservation at -80 °C for 3 months, the minimum inhibitory concentration (MIC) of NX24r was reduced sharply. Mutations and changes of transcriptome were analyzed after amoxicillin screening and cryopreservation. Mutations in PBP1 (I370T, E428K, T556S) and HefC (M337K, L378F, D976V) were detected in NX24r, which may be the main reason for the induced amoxicillin resistance. No mutations were found in PBP1 or HefC after cryopreservation. However, transcriptome analysis showed that down-regulated genes in the cryopreserved clone were significantly enriched in plasma membrane (GO:0005886), including , , , and . These plasma membrane genes are involved in the biosynthesis and transport function of the membrane. The decreased amoxicillin resistance after cryopreservation may be related to the down-regulation of genes involved in membrane structure and transport function.

摘要

一些阿莫西林耐药菌株在冷冻后对阿莫西林的耐药性急剧下降。在中国，大多数临床胃黏膜标本在冷冻后运输，用于幽门螺杆菌的分离和药敏试验，这可能导致对阿莫西林耐药性的低估。本研究的目的是调查冷冻保存后阿莫西林耐药性降低的原因。通过阿莫西林压力筛选获得了一株高水平阿莫西林耐药克隆（NX24r）。在-80℃冷冻保存3个月后，NX24r的最低抑菌浓度（MIC）急剧降低。对阿莫西林筛选和冷冻保存后的突变及转录组变化进行了分析。在NX24r中检测到PBP1（I370T、E428K、T556S）和HefC（M337K、L378F、D976V）的突变，这可能是诱导阿莫西林耐药性的主要原因。冷冻保存后在PBP1或HefC中未发现突变。然而，转录组分析表明，冷冻保存克隆中下调的基因在质膜（GO:0005886）中显著富集，包括[具体基因名称未给出]、[具体基因名称未给出]、[具体基因名称未给出]、[具体基因名称未给出]和[具体基因名称未给出]。这些质膜基因参与膜的生物合成和运输功能。冷冻保存后阿莫西林耐药性降低可能与参与膜结构和运输功能的基因下调有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/cbf8c2cedfd0/pathogens-10-00676-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/b7283ba74a6a/pathogens-10-00676-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/0fd06110c5f2/pathogens-10-00676-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/cbf8c2cedfd0/pathogens-10-00676-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/b7283ba74a6a/pathogens-10-00676-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/0fd06110c5f2/pathogens-10-00676-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec2/8229390/cbf8c2cedfd0/pathogens-10-00676-g003.jpg

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