用羊血代替兔血制作血平板在“链球菌菌落在血平板上的溶血特性”实验中的应用  

Sheep blood agar plate for identifying streptococcus hemolytic properties

作　　者：徐鸣阳 李美玉[2] XU Ming-yang;LI Mei-yu(Experimental Teaching Center,School of Medicine,Sun Yat-sen University,Guangzhou,Guangdong 510080;Experimental Teaching Center,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,Guangdong 510080,China)

机构地区：[1]中山大学医学院医学实验教学中心,广东广州510080 [2]中山大学中山医学院医学实验教学中心,广东广州510080

出　　处：《热带医学杂志》2020年第4期489-491,520,F0004,共5页Journal of Tropical Medicine

基　　金：中山大学本科教学质量工程项目(中大教务【2018】294号);中山大学校级本科教学质量工程项目(59000-31911131)。

摘　　要：目的本实验用脱纤维羊血代替脱纤维兔血制作血平板,并观察其对链球菌溶血现象产生的影响。方法向高压灭菌过的哥伦比亚琼脂培养基中加入6 mL脱纤维羊血或兔血,混匀后,倾注平板。经无菌试验合格后,将链球菌分区划线涂布于平板上,37℃培养24 h,培养后于室温放置1和2 d后(day 2和day 3),观察菌落在血平板上的生长情况和产生溶血情况。结果甲型溶血性链球菌培养24 h后(day 1),在羊血平板上有明显的草绿色溶血环,而兔血平板上并未产生明显的草绿色溶血环;平板置于室温1和2 d后(day 2和day 3),在羊血平板上仍然能观察到草绿色溶血环,而培养在兔血平板上的菌落在day 3时才观察到微弱的α溶血现象。乙型溶血性链球菌培养24 h后(day 1),在羊血平板上已经出现明显的无色透明溶血环,此时兔血平板上也有溶血环,但并不透明;平板置于室温1 d后(day 2),羊血平板上的无色透明溶血环更加明显和透明,此时兔血平板开始出现无色透明溶血环。丙型链球菌培养24 h后(day 1),羊血和兔血平板上,均未产生溶血现象,在室温静置几天后,也没有产生溶血现象。结论羊血平板更适合观察甲、乙、丙型链球菌的溶血现象,更适用于教学示教用。Objective This paper demonstrates how to use blood agar made with defibrinated sheep blood as rabbit blood substitute for identifying streptococcus hemolytic properties.Methods To make blood agar,6 ml defibrinated sheep blood or rabbit blood was added evenly to sterilized Columbia Blood Agar Base to make agar plates and tested to confirm sterilized condition.Streptococcus colonies were streak plated on the agar plate to obtain single colonies and then incubated for 24 hr at37℃.After the incubation,the plates were kept at room temperature for one day(day 2)and two days(day 3).The colony growth and hemolytic properties on blood agar were observed.Results After incubate for 24 hr(day 1),α-hemolytic streptococcus growing on sheep blood agar showed apparent grass green hemolytic ring.Meanwhile,α-hemolytic streptococcus growing on rabbit blood agar did not generate any apparent grass green hemolytic ring.Theα-hemolytic property of some colonies started to appear on rabbit blood agar on day 3.After incubate for 24 hr(day 1),β-hemolytic streptococcus growing on sheep blood agar had already showed apparent colorless and transparent hemolytic ring,while at the same time some hemolytic ring could also be found on rabbit blood agar,but not that transparent.After incubated at room temperature for one day(day 2),more obvious and transparent hemolytic ring could be seen on sheep blood agar,whileβ-hemolytic streptococcus on rabbit blood agar started to generate colorless and transparent hemolytic ring.After incubate for 24 hr(day 1),no hemolytic properties could be found on neither sheep blood agar,nor rabbit blood agar that growingγ-streptococcus.Even after incubated for several days,it still did not generate any hemolytic properties.Conclusion Agar plate made with sheep blood was more applicable for demonstrating hemolytic properties of streptococcus.

关 键 词：甲型溶血性链球菌 乙型溶血性链球菌 丙型链球菌 羊血平板 兔血平板 

分 类 号：R378.12[医药卫生—病原生物学]