Piersimoni C, Scarparo C, Callegaro A, Tosi C P, Nista D, Bornigia S, Scagnelli M, Rigon A, Ruggiero G, Goglio A
Department of Clinical Microbiology, General Hospital Umberto I degrees -Torrette, Ancona, Italy.
J Clin Microbiol. 2001 Feb;39(2):651-7. doi: 10.1128/JCM.39.2.651-657.2001.
The MB/BacT ALERT 3D System (MB/BacT) (Organon Teknika, Boxtel, The Netherlands) is a fully automated, nonradiometric system with a revised antibiotic supplement kit designed for the recovery of mycobacteria from clinical specimens. In a multicenter study, the recovery rate of acid-fast bacilli (AFB) and the mean time to their detection from clinical specimens was determined by using the MB/BacT system. Data were compared to those assessed by the radiometric BACTEC 460 system (B460) and by culture on Löwenstein-Jensen (L-J) solid medium. A total of 2,859 respiratory and extrapulmonary specimens were processed by the N-acetyl-L-cysteine (NALC)-NaOH method using two different concentrations of sodium hydroxide; 1.5% was adopted in study design A (1,766 specimens), and 1.0% was used in study design B (1,093 specimens). The contamination rates for MB/BacT were 4.6% (study design A) and 7.1% (study design B). One hundred seventy-nine mycobacterial isolates were detected by study design A, with 148 Mycobacterium tuberculosis complex (MTB) isolates and 31 nontuberculous mycobacteria (NTM) isolates. Overall recovery rates were 78.8% for MB/BacT (P = 0.0049), 64.2% for L-J (P < 0.0001), and 87.1% for B460, whereas they were 84.5, 70.9, and 91.2%, respectively, for MTB alone. A total of 125 mycobacteria were detected by study design B, with 46 MTB and 79 NTM. Overall recovery rates by the individual systems were 57.6% (P = 0.0002), 56.8% (P = 0.0001), and 80% for MB/BacT, L-J, and B460, respectively, whereas the rates were 91.3, 78.3, and 97.8% for MTB alone. By study design A, the mean times to detection of smear-positive MTB, smear-negative MTB, and NTM were 11.5, 19.9, and 19.6 days, respectively, with the MB/BacT; 8.3, 16.8, and 16.6 days, respectively, with the B460; and 20.6, 32.1, and 27.8 days, respectively, with L-J medium. By study design B, the mean times were 15.1, 26.7, and 26 days with the MB/BacT; 11.7, 21.3, and 24.8 days with the B460; and 20.4, 28.7, and 28.4 days with L-J medium. Identification was attempted by probing (Accuprobe) MB/BacT-positive bottles within the first working day following instrument positive flag. Results were compared to those obtained in the B460 positive vials by the p-nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test (study design A) or by the Accuprobe assay (study design B). About 90% of MTB and 100% of NTM could be identified, showing turnaround times closely related to those obtained by combining B460 and the NAP test or the Accuprobe assay. In conclusion, even though recovery rates were shown to be lower than B460, especially for NTM, and contaminants were somewhat higher, MB/BacT represents a valuable alternative to the radiometric system, especially in those laboratories where disposal of radioactive waste is restricted. Finally, when AFB are cultured in nonradiometric liquid media, our data (detection times and bacterial overgrowth rates) suggest that decontamination with 1.5% NaOH may be more suitable than the standard NALC-NaOH.
MB/BacT ALERT 3D系统(MB/BacT)(奥加农公司,荷兰博克斯泰尔)是一种全自动、非放射性系统,配备经过修订的抗生素补充试剂盒,用于从临床标本中分离分枝杆菌。在一项多中心研究中,使用MB/BacT系统测定了耐酸杆菌(AFB)的回收率及其从临床标本中检测出来的平均时间。将数据与通过放射性BACTEC 460系统(B460)和在罗-琴(L-J)固体培养基上培养所评估的数据进行比较。使用两种不同浓度的氢氧化钠,通过N-乙酰-L-半胱氨酸(NALC)-氢氧化钠法对总共2859份呼吸道和肺外标本进行处理;研究设计A(1766份标本)采用1.5%的氢氧化钠,研究设计B(1093份标本)采用1.0%的氢氧化钠。MB/BacT的污染率在研究设计A中为4.6%,在研究设计B中为7.1%。研究设计A检测到179株分枝杆菌分离株,其中148株为结核分枝杆菌复合群(MTB)分离株,31株为非结核分枝杆菌(NTM)分离株。MB/BacT的总体回收率为78.8%(P = 0.0049),L-J为64.2%(P < 0.0001),B460为87.1%,而仅MTB的回收率分别为84.5%、70.9%和91.2%。研究设计B检测到125株分枝杆菌,其中46株为MTB,79株为NTM。各系统的总体回收率分别为:MB/BacT为57.6%(P = 0.0002),L-J为56.8%(P = 0.0001),B460为80%,而仅MTB的回收率分别为91.3%、78.3%和97.8%。通过研究设计A,MB/BacT检测涂片阳性MTB、涂片阴性MTB和NTM的平均时间分别为11.5天、19.9天和19.6天;B460分别为8.3天、16.8天和16.6天;L-J培养基分别为20.6天、32.1天和27.8天。通过研究设计B,MB/BacT的平均时间为15.1天、26.7天和26天;B460分别为11.7天、21.3天和24.8天;L-J培养基分别为20.4天、28.7天和28.4天。在仪器出现阳性信号后的第一个工作日内,通过对MB/BacT阳性瓶进行探针检测(Accuprobe)来尝试进行鉴定。将结果与通过对硝基-α-乙酰氨基-β-羟基苯丙酮(NAP)试验(研究设计A)或Accuprobe检测法(研究设计B)在B460阳性小瓶中获得的结果进行比较。约90%的MTB和100%的NTM能够得到鉴定,显示出周转时间与通过结合B460和NAP试验或Accuprobe检测法所获得的周转时间密切相关。总之,尽管回收率显示低于B460,尤其是对于NTM,并且污染物略多,但MB/BacT是放射性系统的一种有价值的替代方法,特别是在那些放射性废物处理受到限制的实验室中。最后,当在非放射性液体培养基中培养AFB时,我们的数据(检测时间和细菌过度生长率)表明,用1.5%氢氧化钠进行净化可能比标准的NALC-氢氧化钠更合适。