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ʱ䣺2015/2/10 13:07:50

Ƽ1 , Ӧ1 , ´Ԫ1 , 2

ժҪĿġ2007 ѮѮijЬбвԭѧоɼȻߵı27 ڻѪ걾17 ݺͻָڻѪ걾12 27 ݱӱ걾,ӫⶨRT - PCR MDCKϸ߷вԭѧѪ걾ѪѧѪ27 ݱӱ걾ӫⶨRT - PCR 17 Ϊ62196%

(17 /27) ; 𷨼1244144% (12 /27) ; MDCKϸ2ݱӱ걾з2H3N2 в7141% (2 /27) ; ߷δв˵˫ѪѧʾָڿȽϼڿۡH3N2 вӫⶨPCR ԸȺвĿټ

ؼʡвӫⶨRT - PCR; ϸ

Etiolog ic Investiga tion of an Outbreak of Influenza - like Illness and Compar ison of the MethodsGAN L i - pingCHEN Ying - jian, CHEN Chun - yuan, et al 13Shenzhen M unicipal Longgang Center for D isease Control and Prevention,Guangdong 518172, China

AbstractObjectiveTo exp lore the pathogen of an outbreak of influenza - like illness among workers of a shoefactory in Longgang district, Shenzhen city in 20071MethodsThe pathogens in 27 nasopharyngeal samp les from patientswithcute fever were detected by using fluorescent quantitative multip lex PCR, colloidal gold method, MDCK cell culture and

chicken embryo cell culture1Hemagglutination inhibition test and serological subtyp ing method were app lied for analyzing 29serum samp les obtained from 17 patients in acute phase and 12 patients in convalescent period1ResultsThe positive rates ofthe swab samp leswere 62192% (17 /27) , 44144% (2 /27) and 7141% (2 /27) , respectively by FQ - PCR, MDCK cell culture and colloidal gold method1No influenza viruswas detected in chicken - embryo culture1Analysis of paired serum samp les from 6patients revealed 4 - fold or more antibody increase in convalescent period compared with acute phase1Conclusion This outbreak was caused by influenza H3N2 virus1 FQ - PCR is a rap id, specific and sensitive method for the detection of influenza

virus1

Key wordsInfluenza virus; FQ - PCR; Colloidal gold method; Cellular culture



вԷпȫʹΪҪ֢״ļϺȾȾڹѧУȺȺеĵط𱩷2007 Ѯ6ѮijЬһȺԷȵ߳¡38пȫʹ֢״Ϊ˽ⲡɼȻ߱ӺѪвԭѧ÷ѧʹͳϵķȷϸΪH3N2

вϺȾ

1Ϻͷ

11          

111걾IJɼʹ걾Դ2007ѮѮijЬԭȺԷȻɼߵı27 , 2 h ʵҼ2 000 U /ml ù2 000 U /mlùغ100 U /mlճB, 4汸

24 hϸ뼦ߵ70ͬʱɼڻѪ걾17 ݺͻָڻѪ걾12 

112ȡԼ: Roche High Pure Viral RNAKitڵ¹Qiagen˾

113A /B вӫPCR ԼпƷ޹˾

114ϸ: MDCKϸΪмԤṩ; MEMСţѪΪGib2co /BRL ˾11115 ׼Ѫ: A /panama /2007 /99 (H3N2 ) WHO ṩ

116Ըð𷨿Լɺݴؼ޹˾ṩ

12AB IӦϵͳ˾7500 RealTime PCR ӫⶨPCR 

13

131ӫⶨRT - PCR 

1311ȡQiagen ˾RNA Լ˵вRNA ij

1312ӫⶨRT - PCRӦ: AӫⶨRT - PCR Լ˵

:(1) PCR ӦϵӦ25l, ں8ģ, 16125ӦҺ, 015l Taq ø,0125lת¼ø

(2) PCR Ӧ42 30 min ת¼952 min, 95 5 s, 60 1 min 40 ѭ60ӫӫѡѡFAM ROX˫ɫӫͨ

(3) AB I 7500 Real Time PCR Ǽ˻׶εӫ

(4) ж:ԶCt3010,ԶCt3010,ƷCt3010,Ʒ

132Լ˵

133ϸMDCKϸ뷨ѳɵMDCKϸȥҺHanksҺϴÿݱ걾ÿֱܽ걾011 ml, 351 h 30 min, HanksҺϴ21 mlţѪάҺ,35

쿪ʼ۲ϸ(CPU) , 80% CPU ʱ΢Ѫ(H I) , Ա͡1

13GB15994 - 1995 ı׼2

135кѪѪ1ϡ1256ͬʱⶨںͻָѪ岡,ԻָѪ忹ζȱȼ4Ϊ

2

21ӫⶨRT - PCR A /B ӫⶨRT - PCR , 27 ݱ, 17 вΪ

62196% (17 /27) ; B вԽ27ݱ걾߼ͼ1, жCT ֵ1

22𷨼

27 ݱӱ걾нAн𷨵Ŀټ12 44144%(12 /27) ; B вԭԷӦΪвԸð

23кѪ

ڲѪ17 ָڲѪ12 ,˫Ѫѧʾ˫Ѫ, AвѪָڿȽϼڿ

24MDCKϸ߷

ӫⶨRT - PCR Ա걾ͬʱMDCKϸͼ߷ݱӱ걾ҺвA /B ͺͱ׼ѪH IҺH3N2 ׼ѪѪͻ͵ı׼ѪδH3N2 в7141% (2 /27) ; ߷δ뵽в215ӫⶨRT - PCRѪѧ2

3֡

2007ѮѮijЬԭȺԷó8ֳ1Ź17 6024 93127 ,5510 /Ů12 671 188 148100 /ͳѧ(2 = 25178, P < 010001) , ŮʲͳѧŮԷԶԷŮְռϴ71199 % (12 671 /17 602) й鹲215 ˷,Ϊ122115 /Ҫٴ֢״Ϊ38ʹͷʹȫ֫ʹ״ְб֢״Ķٴ֢״,30ն10֢Ӳֳн𷨿ټвRT - PCR7Aв1 ٴβɼ֢17ֳ

𷨿ټ7AвRT -PCR10AввѧѪѧѧ֤֤ʵһH3N2 вľֲ

27 걾ӫⶨRT - PCR Ϊ62196% ( 17 /27) , 𷨼Ϊ44144% (12 /27) , ϸΪ7141% ( 2 /27) ߷12 ˫ѪݻָڿȽϼڿӼۺϷӫⶨRT - PCR Ը뱨صӫⶨRT -PCR ԸMDCKϸһ¡2, 3ΪӫⶨRT - PCR dzǿԭʼ걾мʹٵIJҲܼ⵽в34 hڵó,ΪٵвϷ𷨵ļײֳ,ǿȽӫⶨRT - PCR ѪѧҪɼ˫ѪʱȽϳҲײɵָڵѪΪԱԽϴӰؽ϶ٵı걾һϸ߷뵽ҼϸԺͲɼı걾΢ȾӰ˲ķ롲4ϸķʵ,걾IJҪ󼰱걾ɼҪϸйء2ӱ걾δ뵽ввԼ߲йӫⶨRT - PCR ʹͳͬʱٴвѧԿٲ𱾴μIJΪ

ṩ

Ρġ

1GB1594 - 19951Ըðϱ׼ԭ1й׼, 20031131 - 138

2СԽ1ӫⶨRT - PCR вϵӦ1лʵٴѧ־, 2004, 18: 289 -

2901

3ѧŽȺ1ӫⶨRT - PCR12ѧУȺԷȼй־, 2005, 1352 -

13531

4EchavarriaM, FormanM, Ticehurst J, et al1 PCR method forde2tection of adenovirus in urine of healthy and human immunodeficien- cy virus - infected individuals1 J CliniMicrobiology, 1998, 36:3323 - 33261