Madhavan Hajib N, Therese Kulandai L, Doraiswamy Kavitha
Microbiology Research Centre, Vision Research Foundation, Sankara Nethralaya, 18 College Road, Chennai-600 006.
Indian J Ophthalmol. 2002 Mar;50(1):35-9.
To apply polymerase chain reaction (PCR) on vitreous fluid (VF) from Eales' disease to further confirm its association with Mycobacterium tuberculosis.
Sixty nine VF samples from 69 patients (24 Eales' disease and 45 Non-Eales' as controls) were processed by conventional methods for detection of mycobacteria. Polymerase chain reaction (PCR) specific for IS 6110 and nested PCR (nPCR) using primers coding for MPB 64 gene were applied on all 69 VF. PCR based dot-blot hybridisation was applied on the IS 6110 amplified products of n PCR-positive VFs.
Conventional methods (direct smear and culture) did not detect mycobacteria in any of the 69 VF samples. Five (20.8%) of 24 VF from Eales' and 2 (4.2%) of 45 VF from control patients tested positive for M. tuberculosis DNA by nPCR. This difference was statistically significant (P < 0.05). All 69 VF were negative by PCR for IS 6110. Two VF of Eales' patients positive by nPCR were also positive by DNA probe dot-blot hybridisation for IS 6110.
Detection of M. tuberculosis DNA by PCR in a significant number of VF of Eales' disease patients reemphasizes the association of this bacterium with Eales' disease.
对伊尔斯病患者的玻璃体液(VF)进行聚合酶链反应(PCR),以进一步证实其与结核分枝杆菌的关联。
对69例患者(24例伊尔斯病患者和45例非伊尔斯病患者作为对照)的69份玻璃体液样本采用常规方法检测分枝杆菌。对所有69份玻璃体液样本应用针对IS 6110的聚合酶链反应(PCR)以及使用编码MPB 64基因的引物进行巢式PCR(nPCR)。对nPCR阳性玻璃体液样本的IS 6110扩增产物进行基于PCR的斑点杂交。
常规方法(直接涂片和培养)在69份玻璃体液样本中均未检测到分枝杆菌。24份伊尔斯病患者的玻璃体液中有5份(20.8%)通过nPCR检测结核分枝杆菌DNA呈阳性,45份对照患者的玻璃体液中有2份(4.2%)呈阳性。这种差异具有统计学意义(P<0.05)。所有69份玻璃体液样本针对IS 6110的PCR检测均为阴性。2份nPCR检测呈阳性的伊尔斯病患者的玻璃体液样本经DNA探针斑点杂交检测IS 6110也呈阳性。
通过PCR在大量伊尔斯病患者的玻璃体液中检测到结核分枝杆菌DNA,再次强调了该细菌与伊尔斯病的关联。