Direkt mikroskopi ile etken saptanamayan tüberküloz menenjit olgularında, PCR yöntemi ile beyin omurilik sıvısında mycobacterium tuberculosis araştırılması
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Date
1999
Authors
Aydın, Ömür
Journal Title
Journal ISSN
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Publisher
Uludağ Üniversitesi
Abstract
Beyin omurilik sıvısında (BOS), çok az sayıda tüberküloz basilinin bulunması nedeniyle, tüberküloz menenjitin erken tanısında bakteriyolojik tanı metodları yetersiz kalmakta; serolojik testlerle kesin sonuçlar alınamamaktadır. Bu nedenle hızlı tanı metodlarına gereksinim vardır. Tüberküloz tanısında kullanılmaya başlanan Polimeraz Zincir Reaksiyonu- Polymerase Chain Reaction-PCR; materyalde bulunan Mycobacterium tuberculosis DNA'sının çoğaltılmasına dayalı; hızlı, duyarlı ve özgül bir testtir. Aynı gün içinde tüberküloz tanısının konmasını ve aynı zamanda tedaviye geçilmesini sağlar. Çalışmada, 152 hastadan BOS alındı. Çalışma grubunda 22 tüberküloz menenjitli hasta; kontrol grubunda 71 menenjitli, 59 nörolojik bulguları nedeni ile BOS alınan hastalar olmak üzere 130 hasta bulunuyordu. Hastaların tümünün BOS'larından direkt preparat hazırlanarak, M tuberculosis saptamak amacıyla Ehrlich Ziehl-Neelsen yöntemiyle boyandı, Lowenstein-Jensen ve Middlebrook 7H12 besiyerlerine ekildi. Sonra M tuberculosis için özgül, IS6110 insertion sekansının IS-1 ve IS-2 primer çiftleri kullanılarak PCR ile çalışıldı.Tüberküloz menenjitli hastaların 15'i (% 68,1) PCR ile pozitif bulundu. Bunlardan sadece 1 hastanın (% 4,5) BOS ve trakea sıvısında Mycobacterium tuberculosis üredi. Bu hastanın BOS, PCR ile de pozitif idi. Hastaların tümünde direkt preparat negatif idi. Bir'den fazla sayıda BOS alınan hastaların % 66,6'sında tedavinin ilk haftası içinde, % 50'sinde ikinci haftası içinde ve % 16,6'sında üçüncü haftası içinde PCR pozitif bulundu. Kabakulak menenjiti olan bir hastada (% 1) PCR ile yalancı-pozitiflik saptandı. Çalışmada, tüberküloz menenjitli hastalar ile pürülan menenjit, viral menenjit ve nörolojik hastalığı nedeniyle BOS alınan hastalarda, PCR tekniğinin duyarlılığı ve özgüllüğü kıyaslandı. Testin duyarlılığı % 68.1, özgüllüğü % 98.5, pozitif prediktif değeri % 93.7, negatif prediktif değeri % 90.9 bulundu. Sonuç olarak BOS gibi steril örneklerde hızlı ve değerli bir tanı metodu olarak kullanılabileceğine karar verildi.
Investigation of The Presence of Mycobacterium tuberculosis, which can not be Detected by Direct Microscopy, in Cerebrospinal Fluids of Patients with Tuberculous Meningitis by Using PCR Technique Since there are too few numbers of tuberculous bacilli in the cerebrospinal fluid (CSF), bacteriological methods are inadequate and serological methods are unreliable for early diagnosis of tuberculous meningitis. Because of this reason, there are some requirement to use new rapid diagnostic methods. Polymerase chain reaction (PCR) that has begun to utilize at the diagnosis of tuberculosis is a rapid, sensitive and specific test, which is based on amplification of Mycobacterium tuberculosis DNA in clinical samples. This method provides an opportunity to diagnose and begin the specific treatment within same day. In this study, CSF samples collected from 152 patients. The study group included CSF samples taken from 22 patients with tuberculous meningitis. As control group, 130 CSF samples taken from 71 cases with meningitis other than tuberculosis and 59 cases with neurological disorder.CSF samples were examined to detect M tuberculosis by using direct staining with Ehrlich Ziehl-Neelsen method, they were inoculated onto Lowenstein- Jensen and in Middlebrook 7H12 medium. Then, PCR analysis specific for M tuberculosis was performed using IS-1 and IS-2 primers which recognise IS6110 insertion sequences. The specific band for M tuberculosis was detected in the CSF samples of 15 cases (68.1 %) with tuberculous meningitis and only one of them (4.5 %) tuberculous bacilli was isolated from both CSF sample and tracheal aspiration fluid. This patient's CSF sample also gave positive result with PCR. Direct staining of all cases were negative. In the CSF samples of the patients taken at different time points of the therapy, the PCR positivity rate was found to be 66.6 % in the first, 50 % in the second and 16.6 % in the third weeks. A false-positive PCR result is only found in a patient with meningitis caused by mumps virus (1 %). In the study, the sensitivity and specificity of the PCR technique in the CSF samples of the patients with tuberculous meningitis, was compared to that of the patients with purulant meningitis, with viral meningitis or with neurological disorder. The sensitivity, specificity, positive and negative predictive values of the test were found to be 68.1 %, 98.5 %, 93.7 % and 90.9 %, respectively. As a result, this study reveals that PCR is a rapid and a feasibl diagnostic method, which may be use in sterile body fluids such as CSF for the early diagnosis of M tuberculosis.
Investigation of The Presence of Mycobacterium tuberculosis, which can not be Detected by Direct Microscopy, in Cerebrospinal Fluids of Patients with Tuberculous Meningitis by Using PCR Technique Since there are too few numbers of tuberculous bacilli in the cerebrospinal fluid (CSF), bacteriological methods are inadequate and serological methods are unreliable for early diagnosis of tuberculous meningitis. Because of this reason, there are some requirement to use new rapid diagnostic methods. Polymerase chain reaction (PCR) that has begun to utilize at the diagnosis of tuberculosis is a rapid, sensitive and specific test, which is based on amplification of Mycobacterium tuberculosis DNA in clinical samples. This method provides an opportunity to diagnose and begin the specific treatment within same day. In this study, CSF samples collected from 152 patients. The study group included CSF samples taken from 22 patients with tuberculous meningitis. As control group, 130 CSF samples taken from 71 cases with meningitis other than tuberculosis and 59 cases with neurological disorder.CSF samples were examined to detect M tuberculosis by using direct staining with Ehrlich Ziehl-Neelsen method, they were inoculated onto Lowenstein- Jensen and in Middlebrook 7H12 medium. Then, PCR analysis specific for M tuberculosis was performed using IS-1 and IS-2 primers which recognise IS6110 insertion sequences. The specific band for M tuberculosis was detected in the CSF samples of 15 cases (68.1 %) with tuberculous meningitis and only one of them (4.5 %) tuberculous bacilli was isolated from both CSF sample and tracheal aspiration fluid. This patient's CSF sample also gave positive result with PCR. Direct staining of all cases were negative. In the CSF samples of the patients taken at different time points of the therapy, the PCR positivity rate was found to be 66.6 % in the first, 50 % in the second and 16.6 % in the third weeks. A false-positive PCR result is only found in a patient with meningitis caused by mumps virus (1 %). In the study, the sensitivity and specificity of the PCR technique in the CSF samples of the patients with tuberculous meningitis, was compared to that of the patients with purulant meningitis, with viral meningitis or with neurological disorder. The sensitivity, specificity, positive and negative predictive values of the test were found to be 68.1 %, 98.5 %, 93.7 % and 90.9 %, respectively. As a result, this study reveals that PCR is a rapid and a feasibl diagnostic method, which may be use in sterile body fluids such as CSF for the early diagnosis of M tuberculosis.
Description
Keywords
Mycobacterium tuberculosis, Polymerase chain reaction, Polimeraz zincir reaksiyonu, Tüberküloz menenjit, Beyin omurilik sıvısı, Tanı testleri, Tuberculous meningitis, Cerebrospinal fluid, Polymerase chain reaction, Diagnostic tests
Citation
Aydın, Ö. (1999). Direkt mikroskopi ile etken saptanamayan tüberküloz menenjit olgularında, PCR yöntemi ile beyin omurilik sıvısında mycobacterium tuberculosis araştırılması. Yayınlanmamış doktora tezi. Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.