Uddin Mohammad Khaja Mafij, Chowdhury Md Raihan, Ahmed Shahriar, Rahman Md Toufiq, Khatun Razia, van Leth Frank, Banu Sayera
International Centre for Diarrhoeal Disease Research, Bangladesh.
BMC Res Notes. 2013 Jul 25;6:291. doi: 10.1186/1756-0500-6-291.
Sputum smear microscopy is fast and inexpensive technique for detecting tuberculosis (TB) in high incidence areas but has low sensitivity. Physical and chemical sputum processing along with centrifugation have been found to show promise in overcoming this limitation. Our objective was to compare the sensitivity of smear microscopy obtained with smears made directly from respiratory specimens to those from concentrated specimens.
By active screening, 915 TB suspects were identified from Dhaka Central Jail and sputum specimens were aseptically collected. Direct smears were prepared by taking a small portion of the purulent part of the sputum with a sterile loop. The specimens were then processed by a standard N-acetyl-L-cysteine-NaOH digestion-decontamination method to prepare concentrated specimens. Both smears were then air dried, heat fixed, and stained by the Ziehl-Neelsen staining technique. The stained slides were examined under oil immersion and were graded following International Union Against Tuberculosis and Lung Diseases guidelines. All the specimens were inoculated into Lowenstein-Jensen (L-J) media and culture results were considered as gold standard to calculate sensitivity.
Of 915 specimens, 73 (8%) specimens were positive both on direct and concentrated methods, one sample was positive on direct microscopy but was negative on concentrated method. An extra 14 (1.5%) samples were positive on concentrated method which were negative on direct smear. In L-J media 105 specimens were found positive for TB bacilli and of them, 74 (70.5%) and 87 (82.9%) were positive in direct and concentrated smear, respectively. The sensitivity of direct and concentrated smear microscopy was different when using positive culture as the gold standard (71% vs. 83%).
The results showed that concentrated technique increases the sensitivity of microscopy up to 12%. Therefore, the national programs in high TB burden countries may consider incorporating the technique into their guidelines at least in the district and higher level laboratories to improve case finding strategy.
痰涂片显微镜检查是在高发病率地区检测结核病(TB)的快速且廉价的技术,但灵敏度较低。已发现物理和化学痰液处理以及离心在克服这一局限性方面显示出前景。我们的目的是比较直接从呼吸道标本制作的涂片与浓缩标本涂片的显微镜检查灵敏度。
通过主动筛查,从达卡中央监狱识别出915名结核病疑似患者,并无菌采集痰液标本。用无菌接种环取一小部分痰液脓性部分制备直接涂片。然后通过标准的N-乙酰-L-半胱氨酸-氢氧化钠消化去污法处理标本以制备浓缩标本。然后将两种涂片空气干燥、热固定,并采用萋-尼染色技术染色。将染色后的玻片在油镜下检查,并按照国际抗结核和肺部疾病联盟的指南进行分级。所有标本均接种到罗-琴(L-J)培养基中,培养结果被视为计算灵敏度的金标准。
在915份标本中,73份(8%)标本直接涂片和浓缩涂片均为阳性,1份标本直接显微镜检查阳性但浓缩涂片阴性。另外有14份(1.5%)标本浓缩涂片阳性而直接涂片阴性。在L-J培养基中,发现105份标本结核杆菌阳性,其中直接涂片和浓缩涂片分别有74份(70.5%)和87份(82.9%)阳性。以阳性培养作为金标准时,直接涂片和浓缩涂片显微镜检查的灵敏度不同(71%对83%)。
结果表明,浓缩技术可将显微镜检查的灵敏度提高12%。因此,结核病负担高的国家的国家项目可考虑至少在地区及更高级别的实验室将该技术纳入其指南,以改进病例发现策略。