国际病毒学杂志
國際病毒學雜誌
국제병독학잡지
INTERNATIONAL JOURNAL OF VIROLOGY
2015年
3期
153-157
,共5页
严寒秋%霍达%刘白薇%高志勇%李伟红%杜轶威%梁志超%杨扬%陈丽娟
嚴寒鞦%霍達%劉白薇%高誌勇%李偉紅%杜軼威%樑誌超%楊颺%陳麗娟
엄한추%곽체%류백미%고지용%리위홍%두질위%량지초%양양%진려연
牡蛎%诺如病毒%前处理%荧光定量RT-PCR
牡蠣%諾如病毒%前處理%熒光定量RT-PCR
모려%낙여병독%전처리%형광정량RT-PCR
Oyster%Norovirus%Pretreatment%Real time RT-PCR
目的 对牡蛎中诺如病毒检测的三种前处理方法进行比较并探讨应用策略.方法 2014年8月-2015年3月采集160只市售牡蛎,每组5只,共32组样品,取其肠腺组织采用直接处理法(M1)、PEG 8000沉淀法(M2)和蛋白酶K消化-PEG 8000沉淀法(M3)三种方法进行前处理,提取核酸后荧光定量RT-PCR检测诺如病毒核酸.应用串并联法对三种前处理方法的样品阳性率进行分析,利用x2检验比较不同方法的阳性率差异,应用一致性检验比较不同方法的一致性,对荧光RT-PCR所得到的Ct值进行ANOVA检验.结果 M1、M2、M3三种前处理方法诺如病毒阳性率依次为15.63%(5/32)、34.38% (11/32)和37.50% (12/32);M1/M2/M3并联使用的阳性率最高,为50.00% (16/32);其次为M2/M3并联法,阳性率为46.88% (15/32);M1+ M3串联或M1+ M2+ M3串联的阳性率最低,均为9.38% (3/32).M2/M3与M1/M2/M3的阳性率经x2检验比较,差异无统计学意义(P>0.05),两种方法经一致性检验,kappa=0.931(P<0.05),一致性较高.M1、M2、M3三种方法检测阳性样品的Ct值依次为33.44±0.66、33.70±1.88和33.42±2.44.经ANOVA检验,差异无统计学意义(P>0.05).结论 常规监测时推荐使用M2或M3,暴发疫情溯源时可考虑使用M2/M3并联法.
目的 對牡蠣中諾如病毒檢測的三種前處理方法進行比較併探討應用策略.方法 2014年8月-2015年3月採集160隻市售牡蠣,每組5隻,共32組樣品,取其腸腺組織採用直接處理法(M1)、PEG 8000沉澱法(M2)和蛋白酶K消化-PEG 8000沉澱法(M3)三種方法進行前處理,提取覈痠後熒光定量RT-PCR檢測諾如病毒覈痠.應用串併聯法對三種前處理方法的樣品暘性率進行分析,利用x2檢驗比較不同方法的暘性率差異,應用一緻性檢驗比較不同方法的一緻性,對熒光RT-PCR所得到的Ct值進行ANOVA檢驗.結果 M1、M2、M3三種前處理方法諾如病毒暘性率依次為15.63%(5/32)、34.38% (11/32)和37.50% (12/32);M1/M2/M3併聯使用的暘性率最高,為50.00% (16/32);其次為M2/M3併聯法,暘性率為46.88% (15/32);M1+ M3串聯或M1+ M2+ M3串聯的暘性率最低,均為9.38% (3/32).M2/M3與M1/M2/M3的暘性率經x2檢驗比較,差異無統計學意義(P>0.05),兩種方法經一緻性檢驗,kappa=0.931(P<0.05),一緻性較高.M1、M2、M3三種方法檢測暘性樣品的Ct值依次為33.44±0.66、33.70±1.88和33.42±2.44.經ANOVA檢驗,差異無統計學意義(P>0.05).結論 常規鑑測時推薦使用M2或M3,暴髮疫情溯源時可攷慮使用M2/M3併聯法.
목적 대모려중낙여병독검측적삼충전처리방법진행비교병탐토응용책략.방법 2014년8월-2015년3월채집160지시수모려,매조5지,공32조양품,취기장선조직채용직접처리법(M1)、PEG 8000침정법(M2)화단백매K소화-PEG 8000침정법(M3)삼충방법진행전처리,제취핵산후형광정량RT-PCR검측낙여병독핵산.응용천병련법대삼충전처리방법적양품양성솔진행분석,이용x2검험비교불동방법적양성솔차이,응용일치성검험비교불동방법적일치성,대형광RT-PCR소득도적Ct치진행ANOVA검험.결과 M1、M2、M3삼충전처리방법낙여병독양성솔의차위15.63%(5/32)、34.38% (11/32)화37.50% (12/32);M1/M2/M3병련사용적양성솔최고,위50.00% (16/32);기차위M2/M3병련법,양성솔위46.88% (15/32);M1+ M3천련혹M1+ M2+ M3천련적양성솔최저,균위9.38% (3/32).M2/M3여M1/M2/M3적양성솔경x2검험비교,차이무통계학의의(P>0.05),량충방법경일치성검험,kappa=0.931(P<0.05),일치성교고.M1、M2、M3삼충방법검측양성양품적Ct치의차위33.44±0.66、33.70±1.88화33.42±2.44.경ANOVA검험,차이무통계학의의(P>0.05).결론 상규감측시추천사용M2혹M3,폭발역정소원시가고필사용M2/M3병련법.
Objective To compare three pretreatment methods of oyster for the detection of norovirus and to discuss the application strategy.Methods From August 2014 to March 2015,160 oysters were collected and were divided into 32 groups of five.Intestinal glands were pretreated with direct treatment (M1),PEG (polyethylene glycol) 8000 precipitation (M2) and proteinase K digestion-PEG 8000 precipitation (M3).Nucleic acid of norovirus was extracted and detected by real time RT-PCR.The positive rates of three pretreatments were analyzed using parallel test or serial test.The positive rates between different groups were compared by x2 test and consistency test.ANOVA was applied to compare the Ct values in real time RT-PCR.Results The positive rates of norovirus of M1,M2 and M3 were 15.63% (5/32),34.38% (11/32) and 37.50% (12/32).Parallel test of M1/M2/M3 showed the highest positive rate of 50.00% (16/32),followed by parallel test of M2/M3 with a positive rate of 46.88% (15/32).The positive rates in serial tests of M1 + M3 or M1 + M2 + M3 was the minimum of 9.38% (3/32).The positive rates in parallel tests of M2/M3 and M1/M2/M3 showed no significant difference by x2 test (P > 0.05),and these two methods showed the high consistency with kappa =0.931 (P <0.05).The Ct values of M1,M2 and M3 by real time RT-PCR were 33.44 ±0.66,33.70 ± 1.88 and 33.42 ±2.44,and no statistical significance was found using ANOVA (P > 0.05).Conclusions The Pretreatment methods M2 or M3 are suitable for routine surveillance,and the parallel test of M2/M3 is recommended for the tracing of norovirus outbreak.