FilmArray G Panel在腹泻病原体检测中的应用

樊玲芳 张京云 沙丹 管红霞

樊玲芳, 张京云, 沙丹, 管红霞. FilmArray GⅠ Panel在腹泻病原体检测中的应用[J]. 疾病监测, 2022, 37(6): 807-812. doi: 10.3784/jbjc.202104070187
引用本文: 樊玲芳, 张京云, 沙丹, 管红霞. FilmArray G Panel在腹泻病原体检测中的应用[J]. 疾病监测, 2022, 37(6): 807-812. doi: 10.3784/jbjc.202104070187
Fan Lingfang, Zhang Jingyun, Sha Dan, Guan Hongxia. Application of FilmArray GⅠ Panel in the detection of diarrhea pathogens[J]. Disease Surveillance, 2022, 37(6): 807-812. doi: 10.3784/jbjc.202104070187
Citation: Fan Lingfang, Zhang Jingyun, Sha Dan, Guan Hongxia. Application of FilmArray G Panel in the detection of diarrhea pathogens[J]. Disease Surveillance, 2022, 37(6): 807-812. doi: 10.3784/jbjc.202104070187

FilmArray G Panel在腹泻病原体检测中的应用

doi: 10.3784/jbjc.202104070187
基金项目: 无锡市卫健委项目(No. T202017);无锡市科技局项目(No. 锡科社[2020]259号-24)
详细信息
    作者简介:

    樊玲芳,女,江苏省无锡市人,学士,实验师,主要从事微生物检测,Email: 1774482307@qq.com

    通讯作者:

    管红霞,Tel: 0510–82727730,Email:ghx-331@163.com

  • 中图分类号: R211; R378

Application of FilmArray G Panel in the detection of diarrhea pathogens

Funds: This study was supported by Wuxi Health Committee (No. T202017) and Wuxi Technology Bureau (No. [2020]259 -24)
More Information
  • 摘要:   目的  探讨FilmArray Gastrointestinal(GⅠ)Panel在腹泻病原体检测中的应用价值。  方法  收集2016年1-9月无锡市人民医院(含儿童医院)肠道门诊就诊病例粪便标本152份,分别采用FilmArray GⅠ Panel及real-time PCR进行检测,对两种方法检测结果进行比较。  结果  FilmArray GⅠ Panel共检出19种病原体,阳性检出率为75.00%(114/152);real-time PCR共检出17种病原体,阳性检出率为68.42%(104/152)。 两种方法检测结果符合率67.76%(103/152)。 对腺病毒F组40/41、EIEC(Enteroinvasive E. coli , EIEC)、弯曲菌和蓝氏贾第鞭毛虫,检测结果符合率100.00%;对札如病毒、肠集聚性大肠埃希菌(Enteroaggregative E.coli, EAEC)、艰难梭菌、沙门菌和霍乱弧菌,检测结果符合率为98.03%、94.08%、96.71%、94.08%和98.68%,检测结果一致性中等(Kappa值为0.563、0.546、0.651、0.610、0.744);对诺如病毒GⅠ/GⅡ、A组轮状病毒、肠产毒性大肠埃希菌(Enterotoxigenic E. coli, ETEC)、肠致病性大肠埃希菌(Enteropathogenic E.coli, EPEC)、类志贺邻单胞菌和副溶血性弧菌,检测结果符合率为98.03%、99.34%、97.37%、92.76%、99.34%、98.68%,检测结果一致性较好(Kappa值为0.892、0.971、0.890、0.764、0.944、0.902),经χ2检验分析,FilmArray GⅠ Panel对EAEC、ETEC、EPEC、艰难梭菌和沙门菌检出率高于real-time PCR,差异有统计学意义(P<0.05)。  结论  与real-time PCR相比,FilmArray检测具有快速、准确、高通量等特点,可用于突发公共卫生事件的快速检测及腹泻病的快速诊断。
  • 表  1  Real-time PCR 检测目标基因

    Table  1.   Genes targeted by real-time Polymerase Chain Reaction

      病原目标基因 引物及探针序列(5'~3')来源

    诺如病毒GⅠ型
    ORF1–ORF2 junctionCOG1_FCGYTGGATGCGNTTYCATGA1
    COG1_RCTTAGACGCCATCATCATTYAC
    RING1(a)_TPFAM-AGATYGCGATCYCCTGTCCA-TAMARA
    RING1(b)_TPFAM-AGATCGCGGTCTCCTGTCCA-TAMARA

    诺如病毒GⅡ型
    ORF1–ORF2 junctionCOG2_FCARGARBCNATGTTYAGRTGGATGAG1
    COG2_RTCGACGCCATCTTCATTCACA
    RING2_TPFAM-TGGGAGGGCGATCGCAATCT-TAMARA

    星状病毒
    RNA-dependent
    RNA polymerase (ORF1b)
    AV1CCGAGTAGGATCGAGGGT2
    AV2GCTTCTGATTAAATCAATTTTAA
    Av_ProbeFAM-CTTTTCTGTCTCTGTTTAGATTATTTTAATCACC-TAMARA

    腺病毒
    hexonAdeno.fwdGCCACGGTGGGGTTTCTAAACTT3
    Adeno.revGCCCCAGTGGTCTTACATGCACATC
    Adeno.probe1FAM-TGCACCAGACCCGGGCTCAGGTACTCCGA-TAMARA

    札如病毒
    VP1 (major capsid protein)SV_FGCTGTTSCYACTGGTGCA4
    SV_RGGCATCCTGTCRTTCCAAGCA
    SV_ProbeFAM-CCAATCSAATGTCCCTGAGGCAATACGSAA-TAMARA

    A组轮状病毒
    NSP3NVP3_FACCATCTWCACRTRACCCTC5
    NVP3_RGGTCACATAACGCCCC
    NVP3_R1GGTCACATAACGCCCCTATA
    NVP3_ProbeFAM-ATGAGCACAATAGTTAAAAGCTAACACTGTCAA-TAMARA
    EAEC
    aggRFor.CAGCGATACATTAAGACGCCTAAAG6
    Rev.CGTCAGCATCAGCTACAATTATTCC
    EPECeaeAFor.CCGATTCCTCTGGTGACGA6
    Rev.CCACGGTTTATCAAACTGATAACG
    ETECsta2 (STh)For.GCTCAGGATGCTAAACCAG7
    Rev.ACAAGCAGGATTACAACACA
    sta1 (STp)For.CCCTCTTTTAGTCAGTCAACT6
    Rev.CAGTAAAATGTGTTGTTCATATTTTCTG
    eltFor.CGGCAGAGGATGGTTACAG7
    Rev.CTGGGTCTCCTCATTACAAG
    STECstx1For.CTCGACTGCAAAGACGTATG6
    Rev.ATTATCCCCTGTGCCACTATC
    stx2For.AATGCCACGCTTCCCAGAATTG6
    Rev.CCACATCGGTGTCTGTTATTAACC
    EIEC / 志贺菌ipaHFor.ACCATGCTCGCAGAGAAACT8
    Rev.TCAGTACAGCATGCCATGGT

    空肠弯曲菌
    hipOhipO-FCTTGCGGTCATGCTGGACATAC9
    hipO-RAGCACCACCCAAACCCTCTTCA

    结肠弯曲菌
    glyAglyA-FAAACCAAAGCTTATCGTGTGC9
    glyA-RAGTGCAGCAATGTGTGCAATG

    艰难梭菌
    tcdAFor.CCCAATAGAAGATTCAATATTAAGCTT10
    Rev.GGAAGAAAAGAACTTCTGGCTCACTCAGGT
    tcdBFor.GTGTAGCAATGAAAGTCCAAGTTTACGC10
    Rev.CACTTAGCTCTTTGATTGCTGCACCT

    类志贺邻单胞菌
    23S rRNAPS23S-FCTCCGAATACCGTAGAGTGCTATCC11
    PS23S -RCTCCCCTAGCCCAATAACACCTAAA

    沙门菌
    ttrRSBCAsal-stf-2TTTCGCCACATCACGGTAGC7
    sal-str-2CAGGAGATTACAACATGGCTAA

    霍乱弧菌
    ompWFor.TTGCTGCTAACGTTGGCTTTG7
    Rev.CTGCTTTGTAGGTTGCCGTTG
    ProbeFAM-TGCTCAATGATAGCTGGTTCCTCAACGC-BHQ1

    副溶血性弧菌
    toxRFor.CTGGTTGCCTTCTATTGAGCAG7
    Rev.TTCTCATACGAGTGGTTGCTGTC
    ProbeFAM-CGCTACGTTAAGCACCATGCAGAAGACTC-BHQ1
    小肠结肠炎耶尔森氏菌ystAFor.ATCGACACCAATAACCGCTGAG12
    Rev.CCAATCACTACTGACTTCGGCT

    隐孢子虫
    18S rRNAFor.CATGGATAACCGTGGTAAT13
    Rev.TACCCTACCGTCTAAAGCTG

    溶组织内阿米巴
    small-subunit (SSU) rRNAFor.ATTGTCGTGGCATCCTAACTCA14
    Rev.GCGGACGGCTCATTATAACA

    蓝氏贾第鞭毛虫
    heat shock protein (HSP)For.AGGGCTCCGGCATAACTTTCC15
    Rev.GTATCTGTGACCCGTCCGAG
    注:EAEC. 肠集聚性大肠埃希菌Enteroaggregative E. coli; EPEC. 肠致病性大肠埃希菌Enteropathogenic E. coli; ETEC. 肠产毒性大肠埃希菌Enterotoxigenic E. coli; STEC. 产志贺毒素大肠埃希菌Shiga toxin-producing E.coli; EIEC. 肠侵袭性大肠埃希菌Enteroinvasive E. coli
    下载: 导出CSV

    表  2  FilmArray G Panel和real-time PCR腹泻病原体检测结果

    Table  2.   Detection results of diarrhea pathogen by Filmarray G Panel and Real-time PCR

    病原体FilmArray GⅠ Panelreal-time PCR 
    阳性数阳性率(%)阳性数阳性率(%)
    病毒性病原
    腺病毒F组 40/4131.9731.97
    星状病毒42.6310.66
    诺如病毒 GⅠ/GⅡ型1610.53159.87
    A组轮状病毒2013.161912.50
    札如病毒53.2921.32
    致泻性大肠埃希菌
    EAEC159.8763.95
    ETEC2315.131912.50
    EPEC3321.712415.79
    STEC21.3200.00
    EIEC10.6610.66
    其它细菌性病原
    弯曲菌1610.531610.53
    艰难梭菌106.5853.29
    类志贺邻单胞菌106.5895.92
    沙门菌
    1610.5395.92
    霍乱弧菌53.2931.97
    副溶血性弧菌127.89106.58
    肠出血性大肠埃希菌O157
    10.66//
    小肠结肠炎耶尔森氏菌00.0000.00
    寄生虫
    隐孢子虫10.6600.00
    溶组织内阿米巴00.0000.00
    蓝氏甲第鞭毛虫21.3221.32
    环孢子虫00.00//
    注:/. 无此项数据; EAEC. 肠集聚性大肠埃希菌Enteroaggregative E. coli; EPEC. 肠致病性大肠埃希菌Enteropathogenic E. coli; ETEC. 肠产毒性大肠埃希菌Enterotoxigenic E. coli; STEC. 产志贺毒素大肠埃希菌Shiga toxin-producing E.coli; EIEC. 肠侵袭性大肠埃希菌Enteroinvasive E. coli
    下载: 导出CSV

    表  3  FilmArray G Panel和real-time PCR混合感染检测结果

    Table  3.   Detection of mixed infection by FilmArray G Panel和real-time PCR

     感染病原种类FilmArray GⅠ
    Panel (例)
    real-time
    PCR(例)
    2种病原混合感染
    A组轮状病毒/星状病毒11
    A组轮状病毒/诺如病毒GⅠ/GⅡ型10
    A组轮状病毒/沙门菌21
    A组轮状病毒/艰难梭菌42
    A组轮状病毒/隐孢子虫10
    诺如病毒GⅠ/GⅡ型/艰难梭菌10
    诺如病毒GⅠ/GⅡ型/EPEC11
    诺如病毒GⅠ/GⅡ型/EAEC01
    札如病毒/星状病毒10
    札如病毒/副溶血性弧菌11
    EPEC/ETEC75
    EPEC/EAEC02
    EPEC/沙门菌21
    EPEC/艰难梭菌22
    EPEC/霍乱弧菌11
    EPEC/副溶血性弧菌22
    EPEC/类志贺邻单胞菌02
    ETEC/EAEC30
    ETEC/艰难梭菌10
    类志贺邻单胞菌/EAEC11
    类志贺邻单胞菌/副溶血性弧菌10
    类志贺邻单胞菌/沙门菌11
    弯曲菌/艰难梭菌10
    弯曲菌/蓝氏贾第鞭毛虫11
    弯曲菌/沙门菌11
    3种病原混合感染
    诺如病毒GⅠ/GⅡ型/EPEC/EAEC20
    诺如病毒GⅠ/GⅡ型/STEC/
    副溶血性弧菌
    10
    A组轮状病毒/EPEC/类志贺邻单胞菌01
    星状病毒/EPEC/EAEC20
    札如病毒/STEC/O15710
    札如病毒/EPEC/霍乱弧菌10
    副溶血性弧菌/
    类志贺邻单胞菌/蓝氏贾第鞭毛虫
    01
    EPEC/EAEC/弯曲菌11
    EPEC/EAEC/类志贺邻单胞菌10
    EPEC/ETEC/沙门菌10
    EPEC/ETEC/类志贺邻单胞菌10
    EPEC/ETEC/艰难梭菌01
    EPEC/ETEC/EAEC21
    EPEC/类志贺邻单胞菌/
    副溶血性弧菌
    11
    ETEC/类志贺邻单胞菌/
    副溶血性弧菌
    11
    弯曲菌/沙门菌/
    副溶血性弧菌
    10
    类志贺邻单胞菌/沙门菌/
    霍乱弧菌
    10
    4种病原混合感染
    EPEC/EAEC/类志贺邻单胞菌/
    A组轮状病毒
    10
    EPEC/ETEC/沙门菌/
    艰难梭菌
    10
    EPEC/副溶血性弧菌/
    蓝氏贾第鞭毛虫/类志贺邻单胞菌
    10
      合计5733
    注:EAEC. 肠集聚性大肠埃希菌Enteroaggregative E. coli; EPEC. 肠致病性大肠埃希菌Enteropathogenic E. coli; ETEC. 肠产毒性大肠埃希菌Enterotoxigenic E. coli; STEC. 产志贺毒素大肠埃希菌Shiga toxin-producing E.coli; EIEC. 肠侵袭性大肠埃希菌Enteroinvasive E. coli
    下载: 导出CSV

    表  4  FilmArray G Panel和real-time PCR每种病原检测结果比较

    Table  4.   Comparison of FilmArray G Panel and real-time PCR for detection of each pathogen

      病原体FilmArray
    GⅠ Panel
    real-time PCR阳性符合率
    (%)
    阴性符合率
    (%)
    总体符合率
    (%)
    Kappa值χ2P
    +
    腺病毒F组40/41+ 3 0 100.00 100.00 100.00 1.000//
    0149
    星状病毒+13100.0098.0198.030.3943.0000.083
    0148
    诺如病毒GⅠ/GⅡ型+14293.3398.5498.030.8920.330 0.564
    1135
    A组轮状病毒+191100.0099.2599.340.9711.0000.317
    0132
    札如病毒+23100.0098.0098.030.5633.0000.083
    0147
    EAEC+69100.0093.8494.080.5469.000 0.003
    0137
    ETEC+194100.0096.9997.370.8904.000 0.046
    0129
    EPEC+231095.8392.1992.760.7647.360 0.007
    1118
    STEC+02/98.6898.680.0002.0000.157
    0150
    EIEC+10100.00100.00100.001.000//
    0151
    弯曲菌+160100.00100.00100.001.000//
    0136
    艰难梭菌+55100.0096.6096.710.6515.0000.025
    0142
    类志贺邻单胞菌+91100.00 99.3099.340.9441.000 0.317
    0142
    沙门菌+8888.8994.4194.080.6105.440 0.020
    1135
    霍乱弧菌+32100.0098.6698.680.7442.000 0.157
    0147
    副溶血性弧菌+102100.0098.5998.68 0.9022.000 0.157
    0140
    隐孢子虫+01/99.3499.34 0.0001.000 0.317
    0151
    蓝氏贾第鞭毛虫+20100.00100.00100.001.000//
    0150
    注:+. 阳性; –. 阴性; /. 无此项数据; EAEC. 肠集聚性大肠埃希菌Enteroaggregative E. coli; EPEC. 肠致病性大肠埃希菌Enteropathogenic E. coli; ETEC. 肠产毒性大肠埃希菌Enterotoxigenic E. coli; STEC. 产志贺毒素大肠埃希菌Shiga toxin-producing E.coli; EIEC. 肠侵袭性大肠埃希菌Enteroinvasive E. coli
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-04-07
  • 录用日期:  2022-02-08
  • 网络出版日期:  2022-02-08
  • 刊出日期:  2022-07-14

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