脑脊液抗酸瑞吉双重染色法的建立与评价

Establishment and evaluation of acid fast-Wright’s double staining in cerebrospinal fluid

目的 建立脑脊液抗酸瑞吉双重染色法,并评价其诊断结核性脑膜炎的价值。方法 建立抗酸瑞吉双重染色法:脑脊液用吸管轻轻吹打混匀,取0.5 mL加入Cytospin-4型甩片机中,采用28×g离心5 min;待玻片干燥后,滴加碱性复红溶液(碱性复红8 g溶于10 mL乙醇中,加5%石碳酸900 mL,放置24 h后过滤备用),染色10 min;再用5%盐酸-75%乙醇脱色1 min;然后滴加瑞氏吉姆萨A液约0.5 mL到涂片上,最后将B液加于A液上面,使A、B染液充分混匀,染色5 min;水洗、干燥、镜检。选取河北省胸科医院2018年10月—2019年10月高度疑似结核性脑膜炎的患者48例,采集治疗前脑脊液分别进行抗酸瑞吉双重染色、离心涂片抗酸染色和改良抗酸染色,以临床诊断为金标准,计算3种方法在结核性脑膜炎诊断中的灵敏度、特异度、阳性预测值、阴性预测值、一致率、Kappa值。结果 抗酸瑞吉双重染色的灵敏度为81.25%,特异度为100.00%,阳性预测值为100.00%,阴性预测值为72.73%,一致率为87.50%,Kappa值为0.74;离心涂片抗酸染色的灵敏度为6.25%,特异度为100.00%,阳性预测值为100.00%,阴性预测值34.78%,一致率为37.50%,Kappa值为0.04;改良抗酸染色的灵敏度为87.50%,特异度为100.00%,阳性预测值为100.00%,阴性预测值为80.00%,一致率为91.67%,Kappa值为0.82。结论 抗酸瑞吉双重染色对结核性脑膜炎诊断的灵敏度、特异度均高,可以节省标本,提高诊断效率,值得推广应用。

Objective tuberculous meningitis.MethodsEstablishment of acid fast-Wright’s double staining: 0.5 mL of CSF specimens were loaded into the chamber in which poly-L-lysine-coated slides were inserted and centrifuged at 28×g for 5 minutes. After the chamber were removed and the slides were dried, basic fuchsin solution(8 g of basic fuchsin was dissolved in 10 mL of ethanol, after which 900 mL of 5% carbolic acid was added and mixed, 24 hours later, the mixture was filtered for use) was dropped onto the slides and standed for 10 minutes, then decolorized with 5% hydrochloric acid-75% ethanol for 1 minutes, finally, 0.5 mL of Wright Giemsa solution A and solution B was added successively and fully mixed on the slides, 5 minutes later, washed the mixed solution and dried the slides before microscopy. 48 patients in Hebei Chest Hospital from October 2018 to October 2019were included in the analysis. The cerebrospinal fluid were collected and detected by acid fast-Wright’s double staining, antiacid staining and modified Ziehl-Neelsen stain. Clinical diagnosis as the gold standard and sensitivity, specific, positive and negative predictive value, concordance rate and the Kappa value of the three methods were calculated.Results sensitivity of acid fast-Wright’s double staining was 81.25%, specificity 100.00%, positive predictive value 100.00%, negative predictive value 72.73%, concordance rate 87.50%, and the Kappa value 0.74. The sensitivity, specificity, positive predictive value, and the negative predictive value of anti-acid staining were 6.25%, 100.00%, 100.00% and 34.78%, respectively, the concordance rate 37.50%, and the Kappa value 0.04. The sensitivity of modified Ziehl-Neelsen stain was 87.50%, specificity100.00%, positive predictive value 100.00%, negative predictive value 80.00%, the concordance rate 91.67%, and the Kappa value 0.82.Conclusions and also helps to save enough samples for other tests. It is worthy of widespread promotion in the laboratories.