CAJ | 학술논문

目的探讨脐血采分储存中细菌污染的种类、途径和规律性以及深低温冻存对细菌活力的影响.方法在2000年至2007年脐血干细胞采分常规工作中,留取有核细胞分离后血浆-红细胞悬液10ml,分别注入需氧和厌氧细菌培养瓶,用BacT/ALERT 3D-480全自动血液培养系统培养7d,细菌培养阳性者作废弃处理.同时选取细菌阳性的新鲜脐血87份进一步培养24h,获得纯培养后,将革兰阳(阴)性菌鉴定卡放入全自动微生物分析系统读数孵育器,分别进行需氧菌与厌氧菌鉴定.另外,为观察深低温冻存对细菌的影响,取出冻存6～7年的细菌阳性脐血96份,37℃速融后,取有核细胞浓缩物10 ml,用上述方法做二次细菌培养检测.结果 2000年至2007年采集脐血19 062份,细菌培养阳性336份,细菌污染率为1.8％；作细菌鉴定87份,发现兼性生长58份(67％),专性需氧生长38份(43.7％),专性厌氧生长17份(19.5％)；革兰阴性菌占68％,革兰阳性菌占32％.细菌种类:大肠埃希菌最常见,占25.3％,其次为中间链球菌占14.9％,紫色色杆菌占9.2％.96份细菌鉴测阳性并移至液氮保存6～7年的脐血标本中,经复检仍有83份(86％)保持细菌活性.结论 2000年至2007年脐血采分过程中细菌污染率为1.8％,污染菌在液氮冻存6～7年后86％细菌仍存活.故在产房采集脐血时应加强无菌措施,在临床使用冻存干细胞时应加强细菌复检.
목적 탐토제혈채분저존중세균오염적충류、도경화규률성이급심저온동존대세균활력적영향.방법 재2000년지2007년제혈간세포채분상규공작중,류취유핵세포분리후혈장-홍세포현액10ml,분별주입수양화염양세균배양병,용BacT/ALERT 3D-480전자동혈액배양계통배양7d,세균배양양성자작폐기처리.동시선취세균양성적신선제혈87빈진일보배양24h,획득순배양후,장혁란양(음)성균감정잡방입전자동미생물분석계통독수부육기,분별진행수양균여염양균감정.령외,위관찰심저온동존대세균적영향,취출동존6～7년적세균양성제혈96빈,37℃속융후,취유핵세포농축물10 ml,용상술방법주이차세균배양검측.결과 2000년지2007년채집제혈19 062빈,세균배양양성336빈,세균오염솔위1.8％；작세균감정87빈,발현겸성생장58빈(67％),전성수양생장38빈(43.7％),전성염양생장17빈(19.5％)；혁란음성균점68％,혁란양성균점32％.세균충류:대장애희균최상견,점25.3％,기차위중간련구균점14.9％,자색색간균점9.2％.96빈세균감측양성병이지액담보존6～7년적제혈표본중,경복검잉유83빈(86％)보지세균활성.결론 2000년지2007년제혈채분과정중세균오염솔위1.8％,오염균재액담동존6～7년후86％세균잉존활.고재산방채집제혈시응가강무균조시,재림상사용동존간세포시응가강세균복검.
Objective To evaluate bacteria contamination during collection,processing and storage of cord blood to gain insight into contamination mechanism and direct prevention.Methods Fresh cord blood was separated by hydroxyethyl starch (HES) to harvest nucleated cells.The bacteria contamination was tested by culturing 10 ml plasma-red cells with BacT/ALERT 3D-480 automatic blood culture system.Total 87 positive samples were further identified for bacteria species.Ninety six cord blood nucleated cells concentrate with bacteria positive stored in liquid nitrogen(LN2) for 6-7 years were thawed at 37 C and re-cultured for bacteria analysis.Results We collected 19 062 umbilical cord blood.Among them,336 was bacteria positive ( contamination rate 1.8 ％ ).Eighty-seven positive samples were further investigated with facultative bacteria 58 (66.7 ％ ),aerobic 38(43.7％ ) and anaerobic 17( 19.5％ ),Gram- negative accounted for 68％ while positive 32％.The most frequent bacteria were Escherichia coli ( 25.3％ ),Streptacoccus intermediate ( 14.9％ ) and Chromobacteria violaceum(9.2％ ).Ninety-six nucleated cells concentrate with bacteria positive were cryopreserved at liquid nitrogen for researching.Of them,83 samples( 86％ ) showed positive of bacteria culture after deep-low temperature storage for 6-7 years.Conclusions Bacteria contamination rate of the cord blood collection,processing and storage in 2000 ～ 2007 was 1.8％.Stored in liquid nitrogen for 6-7 years,the viability of bacteria was 86％.The aseptic procedures of cord blood collection in delivery room should be intensified.The bacteria re-culture following thawing of cord blood cells is necessary before clinical transfusion.