• Users Online: 1195
  • Home
  • Print this page
  • Email this page


 
 Table of Contents  
ARTICLE
Year : 2012  |  Volume : 1  |  Issue : 4  |  Page : 190-195

Comparisons among the diagnostic methods used for the detection of extra-pulmonary tuberculosis in Bangladesh


1 Department of Microbiology, Stamford University , 51 Siddeswari Road, Dhaka 1217, Bangladesh
2 National Tuberculosis Reference Laboratory (NTRL), National Institute of Diseases of Chest and Hospital (NIDCH), Mohakhali, Dhaka 1212, Bangladesh

Date of Web Publication1-Mar-2017

Correspondence Address:
Rashed Noor
Department of Microbiology, Stamford University Bangladesh, 51 Siddeswari Road, Dhaka 1217
Bangladesh
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.1016/j.ijmyco.2012.10.004

Rights and Permissions
  Abstract 


The present study was an attempt to establish a suitable method for the effective diagnosis of extra-pulmonary tuberculosis in Bangladesh. In this regard, detection of Mycobacterium tuberculosis from 390 different extra-pulmonary specimens was performed by Bright-Field microscopy, light-emitting diode fluorescence microscopy and Lowenstein–Jensen culture methods, followed by an extensive comparison among these methods. M. tuberculosis was detected in 53 cases through the conventional Lowenstein–Jensen culture method; 49 cases were detected under Bright-Field microscope, whereas the light-emitting diode fluorescence microscopy detected 64 cases. Out of 53 culture-positive isolates, 12 were found to be multi-drug resistant. Light-emitting diode fluorescence microscopy was found to be more sensitive and effective than both the Bright-Field microscopy and the Lowenstein–Jensen culture methods. Incidentally, light-emitting diode fluorescence microscopy appeared imperative to detecting the multi-drug resistant tuberculosis.

Keywords: Extra-pulmonary tuberculosis (TB), Light-emitting diode (LED) fluorescence microscopy, Lowenstein–Jensen (L–J) culture


How to cite this article:
Munshi SK, Rahman F, Kamal S M, Noor R. Comparisons among the diagnostic methods used for the detection of extra-pulmonary tuberculosis in Bangladesh. Int J Mycobacteriol 2012;1:190-5

How to cite this URL:
Munshi SK, Rahman F, Kamal S M, Noor R. Comparisons among the diagnostic methods used for the detection of extra-pulmonary tuberculosis in Bangladesh. Int J Mycobacteriol [serial online] 2012 [cited 2022 Aug 11];1:190-5. Available from: https://www.ijmyco.org/text.asp?2012/1/4/190/201250




  Introduction Top


Mycobacterium tuberculosis (MTB), the causative agent of the disease tuberculosis (TB), is of great global epidemic importance. The bacterium affects not only lungs, but also the other parts of the body system which is generally termed as extra-pulmonary tuberculosis [1],[2]. The majority of TB manifestations are pulmonary, with extra-pulmonary TB comprising around 15% of the reported cases, especially among the immunocompromised patients [3]. The disease remains one of the fatal health problems in Bangladesh with 353,103 new cases, including the extra-pulmonary TB cases, every year and 70,000 deaths annually [4]. However, the discrete incidence of the extra-pulmonary TB still remains obscure in Bangladesh owing to the lack of proper diagnosis.

Currently, several diagnostic methods of TB detection are in practice in Bangladesh, among which the Lowenstein–Jensen (L–J) culture and Bright-Field (BF) microscopy are being exercised more frequently [4],[5]. Use of light emitting diode (LED) fluorescence microscope has also been introduced recently in the National Tuberculosis Reference Laboratory (NTRL) in Bangladesh. However, the overall efficacy, including the sensitivity and specificity of the different detection methods for the extra-pulmonary TB diagnosis, has not been compared yet.

Generally, the slow growth of most pathogenic mycobacteria results in the delay in the definitive diagnosis of TB through the culture method [6]. Direct staining for acid-fast bacilli (AFB) has been reported as the most rapid diagnostic method [5]. However, the accuracy of microscopic examination largely depends on the specimen containing a sufficient number of bacteria (>104/ml). Moreover, BF microscopy cannot distinguish M. tuberculosis from atypical mycobacteria [5]. On the contrary, fluorescence microscopy with fluorochrome dyes such as Auramine O or Auramine–rhodamine is known to possess higher degrees of sensitivity and specificity and hence this method is considered as a more accurate test for the diagnosis of TB [7]. Evidently, regarding the diagnostic rapidity and efficacy, the expediency of the detection of mycobacterial DNA in clinical samples by the polymerase chain reaction (PCR)-based techniques is noteworthy [8],[9],[10],[11]. Nevertheless, this method has a very limited use in Bangladesh owing to inadequate logistics and expertise. Therefore, the detection of extra-pulmonary TB in this country largely depends on culture and on BF microscopy, and to a limited extent, on LED fluorescence microscopy [12]. Another facet of TB-related problems in Bangladesh is the continuous heightening of multi-drug resistance (MDR), which commonly develops over the course of improper treatment of the disease [13],[14],[15],[16],[17],[18]. A sporadic survey in 2006 conducted by the Damien Foundation and the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B) showed the overall prevalence of MDR as high as 5.5% [19]. Hence, the management of such a significant rate of MDR-TB needs to be addressed well in the course of TB diagnosis.

Along these lines, the diagnostic efficacy of all three methods of detection of extra-pulmonary TB cases was compared in this study for the first time in Bangladesh. Interestingly, such an assessment also brought a new notion on the detection of MDR-TB.


  Materials and methods Top


Settings

The study was carried out at the National Tuberculosis Reference Laboratory (NTRL), National Institute of Diseases of Chest and Hospital (NIDCH), Bangladesh. NTRL has been certified by Supranational Reference Laboratory (SRL), Antwerp, Belgium.

Ethics approval

The study was approved by the administrative body of NTRL, NIDCH, Bangladesh.

Sample collection

A total of 390 extra-pulmonary specimens, including pus (110), tissue (85), urine (90), ascetic fluid (15), fluid collected after fine needle aspiration cytology (15), gastric lavage (15), cerebrospinal fluid (15), lymph node aspirate (30) and laryngeal swab (15) were tested. Liquid specimens were aseptically collected in a sterile plastic container. Early morning midstream urine was collected in a sterile falcon tube. Gastric lavage sample was collected from an empty stomach. A sterile absorbent cotton swab was used for the collection of laryngeal swab. Transbronchial and other biopsies were taken aseptically and were kept wet during transportation by adding a few drops of sterile 0.9% saline to the tissue.

Sample processing

Tissue samples were homogenized before decontamination [20]; 2–5 ml of all samples except urine was decontaminated by mixing with an equal volume of 4% NaOH. After 15 min, 7mM of phosphate buffer saline (PBS) solution (pH 6.8) was added making the final volume 45 ml. The sample was centrifuged at 3000 g for 15 min, and the pellet was subjected to further analysis [20]. Urine samples were centrifuged at 3000 g for 15 min before decontamination. The pellet was then decontaminated using 0.4% sulfuric acid, neutralized by adding sterile distilled water, and was centrifuged at 3000 g for 15 min [20].

Detection of M. tuberculosis through microscopic methods

Processed specimens were picked using Pasteur pipettes. Smears were air-dried for 15 min and then heat-fixed at 85 °C for 3 min. For Ziehl–Neelsen (Z–N) staining, smears were covered with carbol fuchsin stain, and were heated until the first vapor appeared. After 10 min, smears were washed up, covered with 25% sulfuric acid for 3 min, and 0.1% methylene blue was flooded over them for 1 min. Finally, the washed and dried smears were examined under the BF microscope (Olympus, CX 21) at 1000 × magnification [13],[21]. For Auramine O staining, smears were covered with 0.1% Auramine solution for 15 min, decolorized with 0.5% acid–alcohol for 3 min, and then 0.3% methylene blue was flooded over them for 1 min. After drying, smears were examined under the LED fluorescence microscope (Primostar, Carl Zeiss LED, Germany) at 400 × magnification (455nm) [13],[21],[22].

Detection of M. tuberculosis through L–J culture

Drops (3–4) of the processed sample were introduced onto the slopes of L–J media, incubated at 37 °C and were examined within 3 days for the early recognition of rapidly growing mycobacteria (if present) and/or contamination (if any), followed by the subsequent observation once a week up to 48 days [23]. The final species identification was based on their relatively slow growth rate, appearance of buff colonies, and the characteristic biochemical traits including nitrate reductase activity, catalase activity, and the P-nitrobenzoic acid (PNB) sensitivity [17].

Statistical validation and measurement of microscopic sensitivity and specificity

Data were statistically validated by determining the p values. The sensitivity, specificity, accuracy, positive- and negative-predictive values were also computed to measure the validity of the tests based on true positive, false positive, true negative and false negative results [13],[24]. Thus, the results of different microscopic techniques could be compared with that of the culture method and with each other significantly [25].

Drug susceptibility test (DST)

Culture positive isolates were tested for drug susceptibility patterns by the proportion method [13],[26],[27] against the four commonly used first-line anti-tubercular drugs: streptomycin (SM), isoniazid (INH), rifampicin (RIF) and ethambutol (EMB) at a concentration of 4, 0.2, 40 and 2μg/ml, consecutively. A sterile platinum loop was scraped across the growth along the L–J culture media slope, and was gently shaken over 5–7 sterile glass beads in a tube. After 30 min, aggregates settled at the bottom of the tube, and 2 ml of Tween-80 was added to the homogenous upper part of the supernatant with the similar dimension of the 0.5% MacFarland standard. Then, serial dilutions of the bacterial suspension were prepared with normal saline up to 10−5. L–J media containing the above-mentioned drugs as well as the drug free media (i.e., control) were inoculated with the inoculums from the dilutions 10−3 and 10−5. The relative resistance was estimated using the following formula:



A result of ⩾1 was considered as resistant, while <1 was interpreted as sensitive.


  Results Top


Higher frequency of detection of M. tuberculosis by LED fluorescence microscopy over culture method and BF microscopy

Out of 390 samples, 53 were found to harbor M. tuberculosis detected through the culture method (gold standard), while 64 and 49 samples were found to be positive through LED fluorescence microscopy and BF microscopy, respectively. The positivity was determined by the appearance of relatively small and buff-colored growth on L–J culture media during 4–5 weeks of incubation (Supplement 1A). Positive reactions in nitrate reduction and catalase tests, absence of growth on L–J media containing PNB (500μg/ml) confirmed the culture-positive isolates as the typical M. tuberculosis. The AFB appeared as red, straight or curved rods under the BF microscope (Supplement 1B) and bright yellow or greenish under the LED fluorescence microscope (Supplement 1C). The results were compared side by side and were found to be statistically significant [Table 1]. Notably, the relative positivity through the LED fluorescence microscopy was found 17.19% and 23.44% higher than that of the culture method and the BF microscopy, respectively [Figure 1]. Additionally, as depicted by [Figure 1], the LED fluorescence microscopy appeared to aid in a complete detection of MDR cases over the culture method and the BF microscopy, which is discussed later.
Figure 1: Detection efficacy of extra-pulmonary M. tuberculosis and their multi-drug resistance (MDR) frequency using Lowenstein–Jensen (L–J) culture, LED fluorescence microscopy, and Bright Field (BF) microscopic methods. Striped bars indicate the relative positivity (%). Black bars indicate the detectable MDR cases, and the grey bars are indicative of speculated MDR. The arrowhead indicates the possible risk point of MDR.

Click here to view
Table 1: Comparative detection frequency of different methods (n = 390).

Click here to view


Positivity according to specimen types and method of detection

A two-dimensional analysis of the numbers of positive isolates identified through the three methods of interest along with the specimen types is presented in [Table 2]. Among the extra-pulmonary specimens, the lymph node aspirate samples were found to pose 36.67% positivity in culture. The overall numbers of positive cases were found relatively higher in pus samples than the others. Laryngeal swabs and fluid samples such as ascetic fluid, gastric lavage and cerebrospinal fluid had no positive cases [Table 2].
Table 2: Positivity among different extra-pulmonary specimens.

Click here to view


Diagnostic efficacy of LED fluorescence microscopy

With the specific objective to establish the most efficient method of diagnosis of TB, the diagnostic efficacy was compared among all three methods using culture as the gold standard [Table 3] and [Table 4]. As revealed, the sensitivity of the LED fluorescence microscopy was found significantly higher than that of BF microscopy. However, the specificity, the positive and negative predictive values and the accuracy of both of the microscopic methods did not differ significantly. Overall, the diagnostic efficacy of LED fluorescence microscopy was found far satisfactory compared with that of the BF microscopy as specifically illustrated in [Table 5].
Table 3: Diagnostic efficacy among LED fluorescence microscopy and L–J culture.

Click here to view
Table 4: Diagnostic efficacy between BF microscopy and the L–J culture.

Click here to view
Table 5: Diagnostic efficacy among LED fluorescence microscopy and BF microscopy.

Click here to view


Projection of increased number of MDR cases by LED fluorescence microscopy

As stated earlier, the diagnostic efficacy of the extra-pulmonary TB detection methods also may be revealed through the accuracy of the detection of MDR-TB cases. In this study, 12 (22.64%) out of the 53 culture positive isolates which showed resistance against isoniazid, were also found to be resistant against rifampicin. Eight of them exhibited resistance against streptomycin (15.09%) and 4 showed resistance against ethambutol (7.55%). Hence these isolates appeared to be multi-drug resistant. However, as the LED fluorescence microscopy detected a higher fraction (16.41%) of extra-pulmonary TB cases over the culture method (13.59%); it was assumed that this method could be useful for estimation of the undetected MDR cases by the culture method [Figure 1], and hence might be effective in the assessment of risks to multi-drug resistance. A careful data-interpretation from [Figure 1] gave rise to speculate upon the frequencies of MDR cases to be 27.34% and 20.93% for the LED fluorescence- and BF microscopy positive isolates, respectively. By subtracting the MDR cases found in the culture method (22.64%) from the computed MDR assessed by the LED fluorescence microscopic method (27.34%), the further risk to MDR was found to increase by around 5%.


  Discussion Top


Current methods used for the diagnosis of extra-pulmonary TB in Bangladesh have been found to exhibit relatively low sensitivity in the detection of M. tuberculosis [12]. Various reports around the globe also focused on the similar problem [28],[29],[30]; however, in Bangladesh, a few studies have been conducted in this regard to date. Kamal et al. (2010) investigated the frequency of extra-pulmonary TB only by the culture method, but did not extend the observation to the other detection techniques [12]. This led the current research to broaden the observation of the frequency of M. tuberculosis among different extra-pulmonary specimens by conventional culture, BF microscopy, and by LED fluorescence microscopic techniques, and to further compare their diagnostic efficacies. This is the first report as far as this research goes in Bangladesh on the comparative study of different methods for detecting extra-pulmonary TB.

Although the culture method is considered to be the gold standard for the detection of M. tuberculosis [31],[32], only 13.59% culture-positive cases were detected in this study, while the LED fluorescence microscopy delivered a relatively higher frequency of detection over both the culture method and the BF microscopy. An incidental aspect of this study focused on the growing prevalence of MDR-TB among the extra-pulmonary specimens [33]. As stated earlier, even an apparently revealed assumptive from [Figure 1], the comparatively higher frequency of MDR-TB prevalence in the case of LED fluorescence microscopy positive isolates could be computed. Such a computational approach to assumptive detection of MDR through the LED fluorescence microscopic method would be highly effective in the control of treatment failure cases and hence the overall improvement of the TB situation in Bangladesh.

Currently, the conventional DST is in use for the detection of MDR-TB in Bangladesh; however, it is noteworthy that a new diagnostic tool, namely “GeneXpert MTB/RIF,” has recently been introduced in NTRL for the rapid detection of drug resistance, which could be factually effective in MDR-TB management. Nevertheless, while establishing the method of effective diagnosis of extra-pulmonary tuberculosis in Bangladesh, such an additional finding through the present study might appear interesting in the aid of the overall assessment of MDR-TB cases.

Overall, the findings of this study strongly emphasize the necessity to initiate the use of LED fluorescence microscopy more frequently in Bangladesh for the accurate and rapid detection of extra-pulmonary TB, which could also be used for the risk assessment of MDR-TB, which is not possible to accurately predict by only using the culture method.


  Disclosure Top


The authors have no potential conflict of interests. All authors agreed to the content of the article and contributed significantly: Saurab Kishore Munshi performed the data acquisition, and primarily drafted the article; Farjana Rahman analyzed and interpreted data; S.M. Mostofa Kamal created the concept and design of the study; and Rashed Noor performed the critical revision of the article and approval for submission.


  Acknowledgement Top


We thank the National Tuberculosis Reference Laboratory (NTRL) of NIDCH, Bangladesh, for providing us with the facilities to carry out the experiments.


  Appendix A. Supplementary data Top


Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.ijmyco.2012.10.004.



 
  References Top

1.
A. Fanning, Tuberculosis: 6. Extra-pulmonary disease, CMAJ 160 (11) (1999) 1597–1603.  Back to cited text no. 1
    
2.
S.K. Sharma, A. Mohan, Extrapulmonary tuberculosis, Indian J. Med. Res. 120 (2004) 316–353.  Back to cited text no. 2
    
3.
S.K. Kurup, C. Chan, Mycobacterium-related ocular inflammatory disease: diagnosis and management, Ann. Acad. Med. 35 (3) (2006) 203–209.  Back to cited text no. 3
    
4.
K. Zaman, M. Yunus, S.E. Arifeen, A.H. Baqui, D.A. Sack, S. Hossain, et al, Prevalence of sputum smear-positive tuberculosis in a rural area in Bangladesh, Epidemiol. Infect. 134 (5) (2006) 1052–1059.  Back to cited text no. 4
    
5.
M. Parvez, K.N. Hasan, M. Rumi, S. Ahmed, M. Salimullah, Y. Tahera, et al, PCR can help early diagnosis of pulmonary tuberculosis, Southeast Asian J. Trop. Med. Public Health 34 (1) (2003) 147–153.  Back to cited text no. 5
    
6.
A. Konstantinos, Testing for tuberculosis, Aust. Prescr. 33 (2010) 12–18.  Back to cited text no. 6
    
7.
S. Patino, L. Aamo, M. Cmino, Y. Casart, F. Bartoli, M.J. Garcia, et al, Autofluorescence of mycobacteria as a tool for detection of Mycobacterium tuberculosis, J. Clin. Microbiol. 46 (2008) 3296–3302.  Back to cited text no. 7
    
8.
M. Causse, P. Ruiz, J.B. Gutierrez-Aroca, M. Casal, Comparison of two molecular methods for rapid diagnosis of extrapulmonary tuberculosis, J. Clin. Microbiol. 49 (8) (2011) 3065–3067.  Back to cited text no. 8
    
9.
F. Gomboa, J. Domingue, E. Padila, J.M. Manterola, E. Gazapo, J. Lonca, et al, Rapid diagnosis of extrapulmonary tuberculosis by ligase chain reaction amplification, J. Clin. Microbiol. 36 (5) (1998) 1324–1329.  Back to cited text no. 9
    
10.
G. Mangiapan, M. Vokurka, L. Schouls, J. Cadranel, D. Lecossier, J. van Embden, et al, Sequence capture-PCR improves detection of mycobacterial DNA in clinical specimens, J. Clin. Microbiol. 34 (5) (1996) 1209–1215.  Back to cited text no. 10
    
11.
S. Ortu, P. Molicotti, L.A. Sechi, P. Pirina, F. Saba, C. Vertuccio, et al, Rapid detection and identification of Mycobacterium tuberculosis by real time PCR and Bactec 960 MIGT, New Microbiol. 29 (2006) 75–80.  Back to cited text no. 11
    
12.
S.M.M. Kamal, H.M. Ahsan, S. Ahmed, K.F.M. Ayaz, S. Mahbub, M.S.I. Khan, et al, Isolation and identification of Mycobacterium from extra-pulmonary specimen at NTRL, NIDCH, J. Med. 11 (2010) 128–130.  Back to cited text no. 12
    
13.
R. Noor, S. Akhter, F. Rahman, S.K. Munshi, S.M.M. Kamal, F. Feroz, Frequency of extensively drug resistant tuberculosis (XDR-TB) among re-treatment cases in NIDCH, Dhaka, Bangladesh, J. Infect. Chemother. (2012), http://dx.doi.org/ 10.1007/s10156-012-0490-8.  Back to cited text no. 13
    
14.
R. Iqbal, I. Shabbir, S.U. Khan, S. Saleem, K. Munir, Multidrug resistance tuberculosis in Lahore, Pak. J. Med. Res. 47 (2008) 1.  Back to cited text no. 14
    
15.
B. Mahadev, N. Srikantaramu, P. James, P.G. Mathew, R. Bhagirathi, Comparison between rapid colorimetric mycobacterial isolation and susceptibility testing method and conventional method using LJ medium, Ind. J. Tub. 48 (2010) 129–134.  Back to cited text no. 15
    
16.
R. Sabouni, M. Kourout, I. Chaoui, A. Jordaan, M. Akrim, T.C. Victor, et al, Molecular analysis of multidrug resistant Mycobacterium tuberculosis isolates from Morocco, Ann. Microbiol. 58 (4) (2008) 749–754.  Back to cited text no. 16
    
17.
S.K. Sharma, S. Kumar, P.K. Saha, N. George, S.K. Arora, D. Gupta, et al, Prevalence of multidrug-resistant tuberculosis among category II pulmonary tuberculosis patients, Indian J. Med. Res. 133 (2011) 312–315.  Back to cited text no. 17
    
18.
G. Shiferaw, Y.Woldeamanuel, M. Gebevehu, F. Girmachew, D. Demessie, E. Lemma, Evaluation of microscopic observation drug susceptibility assay for detection of multidrug resistant Mycobacterium tuberculosis, J. Clin. Microbiol. 15 (1) (2007) 1093–1097.  Back to cited text no. 18
    
19.
A.H. Salim, K.J.M. Aung, M.A. Hossain, A. van Deun, Early and rapid microscopy-based diagnosis of the treatment failure and MDR-TB, Int. J. Tuberc. Lung Dis. 10 (11) (2006) 1248–1554.  Back to cited text no. 19
    
20.
P. Monkongdee, K.D. McCarthy, K.P. Chain, T. Tasaneeyapan, N.H. Dung, N.T.N. Lan, et al, Yield of acid-fast smear and mycobacterial culture for tuberculosis diagnosis in people with human immunodeficiency virus, Am. J. Respir. Crit. Care Med. 180 (9) (2009) 903–908.  Back to cited text no. 20
    
21.
I.N. De Kantor, S.J. Kim, T. Friden, A. Lazlo, P.Y. Norvel, H. Reider, et al, Ziehl–Neelsen staining, in: Laboratory services in tuberculosis control-microscopy, second ed., World Health Organization, Geneva, 1998, pp. 27–29.  Back to cited text no. 21
    
22.
Centers for Disease Control and Prevention (CDC), Use of fluorochrome staining for detecting acid-fast mycobacteria, current laboratory practice series, 2000, Available from: <http://wwwn.cdc.gov/dls/lta/TBFluorochrome/ fluorochrome.pdf>, [accessed on January 12, 2012].  Back to cited text no. 22
    
23.
I.N. De Kantor, S.J. Kim, T. Friden, A. Lazlo, P.Y. Norvel, H. Reider, et al, Inoculation and incubation procedure, in: Laboratory Services in Tuberculosis Control – Culture, third ed., World Health Organization, Geneva, 1998, p. 55.  Back to cited text no. 23
    
24.
Kausar MA. Fundamentals of Medical Sciences, second ed., Asian color printing, 2005, pp. 79–81.  Back to cited text no. 24
    
25.
R. Vignesh, P. Balakrishnan, E.M. Shankar, Value of single acid-fast bacilli sputum smears in the diagnosis of tuberculosis in HIV-positive subjects, J. Med. Microbiol. 56 (12) (2007) 1709–1710.  Back to cited text no. 25
    
26.
S.J. Kim, Drug susceptibility testing in tuberculosis: methods and reliability of results, Eur. Respir. J. 25 (2005) 564–569.  Back to cited text no. 26
    
27.
J.C. Palomino, F. Portaels, Simple procedure for drug susceptibility testing of Mycobacterium tuberculosis using a commercial colorimetric assay, Eur. J. Clin. Microbiol. Infect. Dis. 18 (1999) 380–383.  Back to cited text no. 27
    
28.
R.L. Cowie, J.W. Sharpe, Extra-pulmonary tuberculosis: high frequency in the absence of HIV infection, Int. J. Tuberc. Lung Dis. 1 (1997) 159–162.  Back to cited text no. 28
    
29.
I.N. Hayati, Y. Ismail, Y. Zurkurnain, Extrapulmonary tuberculosis: a two-year review of cases at the General Hospital Kota Bharu, Med. J. Malaysia 48 (1993) 416–420.  Back to cited text no. 29
    
30.
K. Noertjojo, C.M. Tam, S.L. Chan, M.M. Chan-Yeung, Extrapulmonary and pulmonary tuberculosis in Hong Kong, Int. J. Tuberc. Lung Dis. 6 (2002) 879–886.  Back to cited text no. 30
    
31.
S. Haldar, S. Chakravorty, M. Bhalla, S.D. Majumder, J.S. Tyagi, Simplified detection of Mycobacterium tuberculosis in sputum using smear microscopy and PCR with molecular beacons, J. Med. Microbiol. 56 (10) (2007) 1356–1362.  Back to cited text no. 31
    
32.
N. Kiraz, L.E.Y. Akgun, N. Kasifoglu, A. Kiremitci, Rapid detection of Mycobacterium tuberculosis from sputum specimens using the FASTPlaqueTB test, Int. J. Tuberc. Lung Dis. 11 (8) (2007) 904–908.  Back to cited text no. 32
    
33.
K.I. Therese, U. Jayanthi, H.N. Madhavan, Application of nested polymerase chain reaction (nPCR) using MPB 64 gene primers to detect Mycobacterium tuberculosis DNA in clinical specimens from extrapulmonary tuberculosis patients, Indian J. Med. Res. 122 (2005) 165–170.  Back to cited text no. 33
    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]


This article has been cited by
1 Emerging diseases in Bangladesh: Current microbiological research perspective
Rashed Noor,Md. Sakil Munna
Tzu Chi Medical Journal. 2015; 27(2): 49
[Pubmed] | [DOI]
2 Molecular Approaches for Detection of the Multi-Drug Resistant Tuberculosis (MDR-TB) in Bangladesh
Tafsina Haque Aurin,Saurab Kishore Munshi,S. M. Mostofa Kamal,Md. Mostafizur Rahman,Md. Shamim Hossain,Thaythayhla Marma,Farjana Rahman,Rashed Noor,Dipankar Chatterji
PLoS ONE. 2014; 9(6): e99810
[Pubmed] | [DOI]
3 Slide drug susceptibility test for the detection of multidrug-resistant tuberculosis in Bangladesh
Rashed Noor,Akter Hossain,Saurab Kishore Munshi,Farjana Rahman,S.M.Mostofa Kamal
Journal of Infection and Chemotherapy. 2013; 19(5): 818
[Pubmed] | [DOI]
4 Evaluation of the effectiveness of BACTEC MGIT 960 for the detection of mycobacteria in Bangladesh
Mehedi Hasan,Saurab Kishore Munshi,Mst. Sabiha Banu Momi,Farjana Rahman,Rashed Noor
International Journal of Mycobacteriology. 2013; 2(4): 214
[Pubmed] | [DOI]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Materials and me...
Results
Discussion
Disclosure
Acknowledgement
Appendix A. Supp...
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed1615    
    Printed62    
    Emailed0    
    PDF Downloaded119    
    Comments [Add]    
    Cited by others 4    

Recommend this journal


[TAG2]
[TAG3]
[TAG4]