【摘要】：目的 了解杭州市第三人民醫(yī)院2008～2014年血液感染細菌和真菌的種類及其耐藥性,探討Verigene-DNA芯片快速檢測血液感染革蘭陽性菌的效果。 方法 采用BacT/ALERT3D血液標(biāo)本培養(yǎng)系統(tǒng)及單側(cè)單瓶、單側(cè)雙瓶和雙側(cè)雙瓶法,分別對臨床送檢19789人份外周血標(biāo)本中細菌和真菌進行分離培養(yǎng),VITEK2Compact系統(tǒng)對獲得的純培養(yǎng)菌株進行鑒定,分別采用VITEK2Compact系統(tǒng)和紙片擴散法檢測各菌株對不同抗生素的敏感性。采用CLSI推薦的紙片擴散法確證試驗檢測細菌分離株p-內(nèi)酰胺酶活性。采用統(tǒng)計學(xué)軟件對不同血培養(yǎng)模式的分離培養(yǎng)陽性率、優(yōu)勢感染細菌或真菌種類、分離菌株藥物敏感性進行分析。另采用Verigene-DNA芯片對分離的98株革蘭陽性菌進行鑒定并檢測其耐藥基因,菌種鑒定結(jié)果與VITEK2Compact系統(tǒng)進行比較,耐藥基因檢測結(jié)果用PCR復(fù)核。 結(jié)果 上述19789外周血標(biāo)本中,雙側(cè)雙瓶法送檢的細菌和真菌血培養(yǎng)陽性率(29,22%)顯著高于單側(cè)單瓶法(8.28%)和單側(cè)雙瓶法(12.34%)(p0.01)。分離的124種1876株細菌中,人葡萄球菌(Staphylococcus hominis)、大腸埃希菌(Escherichia coli)、金黃色葡萄球菌(Staphylococcus aureus)、表皮葡萄球菌(Staphylococcus epidermidis)、溶血葡萄球菌(Staphylococcus haemolyticus)、肺炎克雷伯菌(Klebsiella pneumonia)、頭狀葡萄球菌(Staphylococcus capitis)、屎腸球菌(Enterococcus faecium)、鮑曼不動桿菌(Acinetobacter baumannii)和糞腸球菌(Enterococcus faecalis)分離率為11.46%-3.00%,其余114種細菌分離率均在2.3%以下。分離的18種136株真菌中,近平滑念珠菌(Candida parapsilosis)、白色念珠菌(Candida albicans)、光滑念珠菌(Candida glabrata)和季也蒙念珠菌(Candida guilliermondii)分離率分別為25.00%、21.32%、16.91%和11.76%,其余14種真菌分離率均在8%以下。所有葡萄球菌屬菌株均對利奈唑胺、萬古霉素和替加環(huán)素敏感,但耐甲氧西林凝固酶陰性葡萄球菌(methicillin-resistant CoNS, MRCoNS)檢出率(87.88%,116/132)明顯高于耐甲氧西林金黃色葡萄球菌(methicillin-resistant S. aureus, MRS A)(54.77%,109/199)(p0.01)。92株屎腸球菌和56株糞腸球菌中,替加環(huán)素敏感率均為100%,利奈唑胺敏感率分別為97.01%和95.56%,萬古霉素耐藥腸球菌(vancomycin-resistant enterococcus, VRE)檢出率分別為15.22%(14/92)和8.93%(5/56)。208株大腸埃希菌對亞胺培南、厄他培南、替加環(huán)素、頭孢替坦、阿米卡星、哌拉西林/他唑巴坦和呋喃妥因的敏感率為92.31%-99.52%,但有1.03%和0.48%菌株對碳青霉烯類抗菌藥物厄他培南和亞胺培南耐藥,其中54.31%菌株檢出有β-內(nèi)酰胺酶活性。122株肺炎克雷伯菌對阿米卡星、頭孢替坦、替加環(huán)素和復(fù)方新諾明敏感率為77.05%-79.51%,但有32.71%和31.15%分別對碳青霉烯類抗菌藥物厄他培南和亞胺培南耐藥,其中28.21%菌株檢出有β-內(nèi)酰胺酶活性。71株鮑曼不動桿菌對多粘菌素B、替加環(huán)素、阿米卡星和頭孢哌酮/舒巴坦的敏感率為60%-100%。29株白色念珠菌對氟康唑、伊曲康唑、克霉唑、5-氟胞嘧啶、制霉菌素和兩性霉素B的敏感率均為100%；34株近平滑念珠菌對5-氟胞嘧啶和制霉菌素敏感率均為100%,但對兩性霉素B、伊曲康唑、氟康唑和克霉唑敏感率為85.71%～97.06%；23株光滑念珠菌對5-氟胞嘧啶、兩性霉素B和制霉菌素敏感率均為100%,但對克霉唑、伊曲康唑和氟康唑敏感率為56.52%～85.71%；16株季也蒙念珠菌對5-氟胞嘧啶和兩性霉素B敏感率均為93.75%,但對氟康唑和伊曲康唑敏感率均僅為6.25%。Verigene-DNA芯片對16種98株革蘭陽性菌鑒定結(jié)果與VITEK2Compact系統(tǒng)鑒定結(jié)果總符合率為91.84%(90/98),其中對葡萄球菌屬、腸球菌屬和鏈球菌屬菌株檢測符合率分別為91.67%(55/60),92.59%(25/27)和90.00%(9/10)。Verigene-DNA芯片法鑒定需時約3.1h,但VITEK2Compact系統(tǒng)鑒定需時約43h。22株金黃色葡萄球菌和12株表皮葡萄球菌中,Verigene-DNA芯片法分別檢出21和8株攜帶甲氧西林耐藥相關(guān)mecA基因,PCR檢測結(jié)果與其完全一致；9株糞腸球菌和16株屎腸球菌中,Verigene-DNA芯片法分別檢出1和5株攜帶萬古霉素耐藥相關(guān)vanA基因,PCR檢測結(jié)果與之相同,但PCR檢測結(jié)果顯示有2株屎腸球菌攜帶vanM基因。 結(jié)論 雙側(cè)雙瓶法可提高血標(biāo)本中細菌和真菌分離培養(yǎng)陽性率。優(yōu)勢血液感染細菌依次為人葡萄球菌、大腸埃希菌、金黃色葡萄球菌和表皮葡萄球菌(10%),優(yōu)勢血液感染真菌依次為近平滑念珠菌、白色念珠菌、光滑念珠菌和季也蒙念珠菌(10%)。盡管不同種類的細菌和真菌臨床菌株對不同抗菌藥物的敏感性有差異,但革蘭陽性菌株對利奈唑胺和替加環(huán)素有較高敏感率,革蘭陰性菌株對替加環(huán)素和阿米卡星有較高敏感率,真菌菌株對5-氟胞嘧啶、兩性霉素B和制霉菌素有較高敏感率。與VITEK2Compact系統(tǒng)比較,Verigene-DNA芯片法鑒定血液感染革蘭陽性菌時具有快速、準(zhǔn)確、簡便并能同時檢測細菌耐藥基因的優(yōu)點,值得在臨床實驗室中推廣與應(yīng)用。
[Abstract]:Purpose To understand the species and drug resistance of the blood-infected bacteria and fungi in the third People's Hospital of Hangzhou from 2008 to 2014, and to study the effect of the Veriene-DNA chip on the rapid detection of the Gram-positive bacteria in the blood Fruit. Methods The bacteria and fungi were isolated and cultured in 19789 human peripheral blood samples by using BacT/ ALERT3D blood specimen culture system and one-sided single-bottle, one-sided double-bottle and two-sided double-bottle method. The strains were identified by the VITEK2Compact system and the paper-paper diffusion method, respectively. Sensitivity of the element. The test-tested bacterial isolates, p-, were confirmed using the CLSI-recommended method of paper diffusion. The positive rate of isolation and culture of different blood culture modes, the superiority of the bacteria or the fungi, and the drug sensitivity of the isolated strains were investigated by using the statistical software. Sex analysis was carried out. The isolated 98 strains of Gram-positive bacteria were identified by the Veriene-DNA chip and their drug-resistant genes were tested. The results of the strain identification were compared with the VITEK2Compact system. PC Results In the above 19789 peripheral blood samples, the positive rate of bacterial and fungal blood culture (29,22%) of the two-sided double-bottle method was significantly higher than that of one-sided single-bottle (8.28%) and one-sided double-bottle (12.34 %) (p0.01). Among the 124 isolated strains of 1876, Staphylococcus hominis, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Klebsiella pneumoniae, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Klebsiella pneumoniae, Staphylococcus, Staphylococcus, Staphylococcus, Klebsiella pneumoniae, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Staphylococcus, Klebsiella pneumoniae, Staphyloc@@ S (s capitis), Enterococcus faecium, Acinetobacter baumannii and Enterococcus faecalis were 11.46%-3.00% and the remaining 114 species were isolated. The separation rates of Candida parapsilosis, Candida albicans, Candida glabrata and Candida guillimondii were 25.00%, 21.32%, 16.91% and 11.76%, respectively, and the other 14 fungi were isolated. The isolation rate was less than 8%. All of the strains of Staphylococci were sensitive to rilenalidomide, vancomycin and tigecycline, but the rate of methicillin-resistant coagulase-negative Staphylococci (MRCoNS) (87.88%,116/132) was significantly higher than that of methicillin-resistant S. aureus, MRS A) (54.77%,109/199) (p0.01). The sensitive rate of tigecycline was 100% in 92 strains of Enterococcus faecium and 56 strains of Enterococcus faecium, and the sensitive rate of the rilenalamine was 97.01% and 95.56%, respectively. The detection rates of vancomycin-resistant enterococci (VRE) were 15.22% (14/92) and 8.93% (5/56) respectively. The sensitivity of imipenem, ertapenem, tigecycline, cefotitan, amikacin, zacillin/ tafluvial/ tafluvial and babbitum was 92.31%-99.52%, but 1.03% and 0.48% of the strains were resistant to carbapenem The sensitivity of 122 strains of Klebsiella pneumoniae to amikacin, cefotitan, tigecycline and compound neomycin was 77.05%-79.51%, but 32.71% and 31.15% respectively applied to carbapenem antibacterial drug. The sensitivity of the 71 strains of Acinetobacter baumannii to polymyxin B, tigecycline, amikacin, and ceftriaxone/ sulbactam was 60% to 100%, and 29 strains of Candida albicans were sensitive to fluconazole, Iqu concha, g. The sensitive rate of mycotoxin,5-fluorophonate, nystatin and amphotericin B was 100%, and the sensitivity of 34 strains of near-smooth Candida to 5-fluorophonate and myxin B was 100%. The sensitive rate of the 5-fluorophonate, the amphotericin B and the myxin B was 100%, but the sensitive rate was 56.52% ~ 85.71%, and the sensitive rate was 56.52% ~ 85.71%. The 16 strains were also sensitive to 5-fluoro-cytomycin and amphotericin B. The results of the identification of 16 98 strains of Gram-positive bacteria and VITEK2Compact system were 91.84% (90/98), 91.67% (55/60), 92.59% (25/27) and 90.00% (9/10), respectively. At the time of the identification, about 3.1 h, but when the VITEK2Compact system was identified for about 43 h.22 strains of S. aureus and 12 S. epidermidis, the Verigene-DNA chip method was used to detect 21 and 8 strains of methicillin-resistant related mecA gene, and the result of the PCR was completely consistent with that of the 9 strains of Enterococcus faecalis. And 1 and 5 strains of vancomycin-resistant vanA gene were detected by the Veriene-DNA chip method, and the results of PCR were the same, but the results of PCR showed that 2 strains of enterococcus faecium were found. bacteria-carrying Conclusion The two-sided double-bottle method can increase the blood sample. The positive rate of bacterial and fungal isolates was positive. The dominant blood-infected bacteria were Staphylococcus, E. coli, S. aureus and Staphylococcus epidermidis (10%), and the dominant blood-infected fungi were Candida, Candida albicans, and Nostoglobus glabrata. In spite of the differences in the sensitivity of different kinds of bacteria and fungal clinical strains to different antibacterial drugs, the Gram-positive strain has a high sensitivity to the tigecycline and tigecycline, and the Gram-negative strains are highly sensitive to tigecycline and Amicka. 5-fluoro-1-(5-fluoro-1)-(2-)-(2-)-(5-fluorophenyl) in comparison with that VITEK2compact system, the Veriene-DNA chip method has the advantages of fast, accurate, simple and convenient and simultaneous detection of the bacterial drug resistance gene,
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.5

【參考文獻】

相關(guān)期刊論文 前10條

1 丁爽;閆玲;李若倩;黃平;馬萍;;PCT和CRP對菌血癥的診斷價值比較[J];國際檢驗醫(yī)學(xué)雜志;2014年07期

2 嚴(yán)育忠;范惠清;徐英;周秀梅;陸燕春;;兩種檢測超廣譜β-內(nèi)酰胺酶表型方法的比較[J];中國感染控制雜志;2012年05期

3 李光輝;朱德妹;汪復(fù);倪語星;孫景勇;徐英春;張小江;胡云建;艾效曼;俞云松;林潔;孫自鏞;陳中舉;賈蓓;黃文祥;卓超;蘇丹虹;魏蓮花;吳玲;張朝霞;季萍;王傳清;王愛敏;張泓;孔菁;徐元宏;沈繼錄;單斌;杜艷;楊青;;2011年中國CHINET血培養(yǎng)臨床分離菌的分布及耐藥性[J];中國感染與化療雜志;2013年04期

4 張慧;楊啟文;徐英春;謝秀麗;陳民鈞;;北京協(xié)和醫(yī)院2000~2013年血培養(yǎng)病原菌分布及耐藥性分析[J];檢驗醫(yī)學(xué)與臨床;2014年18期

5 李光輝;朱德妹;汪復(fù);胡志東;李全;孫自鏞;陳中舉;徐英春;張小江;王傳清;王愛敏;倪語星;孫景勇;褚云卓;俞云松;林潔;徐元宏;沈繼錄;蘇丹虹;卓超;魏蓮花;吳玲;張朝霞;季萍;張泓;孔菁;胡云建;艾效曼;單斌;杜艷;;2012年中國CHINET血培養(yǎng)臨床分離菌的分布及耐藥性[J];中國感染與化療雜志;2014年06期

6 徐英春;臨床微生物學(xué)血培養(yǎng)操作規(guī)范[J];中華檢驗醫(yī)學(xué)雜志;2004年02期

7 彭佳;府偉靈;張曉兵;;血培養(yǎng)中真菌的分布及耐藥性分析[J];中華醫(yī)院感染學(xué)雜志;2006年11期

8 李冰;郭珊;呂錦琪;;血流感中的病原菌分布及耐藥性分析[J];中國微生態(tài)學(xué)雜志;2014年03期

9 張凱;喻華;黃湘寧;劉鑫;黃影;;四川省細菌耐藥監(jiān)測網(wǎng)2011～2012年血流感染病原菌分布及耐藥分析[J];實用醫(yī)院臨床雜志;2014年06期

10 楊兵,陳小青,嚴(yán)杰;腹膜炎患者臨床標(biāo)本中無芽孢厭氧菌的分離和鑒定[J];微生物學(xué)雜志;2001年04期

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