\nTuberculosis (TB) is one of the major causes of death in the world which is caused by Mycobacterium\u00a0tuberculosis. The rate of mortality is high if it is conjugated with HIV. The multiple drug resistant strains make\u00a0it difficult to diagnose and treat the disease. It can be easily spread around the world so there is a need to evolve\u00a0new diagnostic methods that are rapid, cost effective and easy to use. Many methods have been used since years\u00a0for diagnosis of disease. Some of the conventional methods are smear microscopy, chest radiography,\u00a0cultivation of mycobacterium. But, in recent years many new molecular and advanced techniques have been\u00a0used that are more specific and sensitive. These techniques are less time consuming and gives very few false\u00a0positive results thereby, making diagnosis cost effective. In this article, some of the conventional and new\u00a0advanced molecular diagnosis methods have been reviewed and there sensitivity and specificity has been taken\u00a0into consideration.
\nKeywords<\/strong>: Diagnosis, tuberculosis, sensitivity.<\/p>\n
INTRODUCTION<\/strong>: The person infected with HIV has high risk of TB.\u00a0The emergence of drug resistant species has made\u00a0it more difficult to diagnose and cure TB. Even\u00a0today in India, two deaths occur every three\u00a0minutes from TB (WHO Report, 2015). One\u00a0woman on every 2.4 men is diagnosed with\u00a0tuberculosis in India. India carries the burden of\u00a030% tuberculosis population of the world\u00a0(McArthur et al., 2016). The prevalence of\u00a0Diabetes Mellitus in tuberculosis patients is high in\u00a0South Eastern Amhara region (Workneh et al.,\u00a02016). Currently, more than 50 diagnostic methods\u00a0are in development stage for the diagnosis of\u00a0tuberculosis (Lienhardt et al., 2016).\u00a0General Characteristics of mycobacterium\u00a0tuberculosis:\u00a0Mycobacterium tuberculosis is a large rod shaped\u00a0non motile bacterium which is distantly related to\u00a0the Actinomycetes. The length of these rods is 2-4\u00a0micrometers and width is 0.2-0.5 um. It is obligate\u00a0aerobic in nature. For this reason, uberculosis is\u00a0mainly found in lungs lobes where full aeration is\u00a0provided. The bacterium has a slow generation time\u00a0of around 15-20 hours.\u00a0It cannot be classified as either gram positive or\u00a0gram negative as it does not show any of the\u00a0characteristics .But it contain murein in the cell\u00a0wall so stains very weakly or not all with gram\u00a0staining (cells referred to as “ghosts”).These are\u00a0stained by carbol fuchsin stain and are referred to\u00a0as acid fast bacilli.<\/p>\n Cell Wall Structure<\/strong><\/p>\n The cell wall structure of Mycobacterium\u00a0tuberculosis is a major determinant of virulence for\u00a0the bacterium. The cell wall complex\u00a0contains peptidoglycan. More than 60% of the\u00a0mycobacterial cell wall is made up of lipid. The\u00a0lipid portion of cell wall comprises of three major\u00a0components, wax-D, mycolic acids and cord\u00a0factors. The high concentration of lipids in the cell wall\u00a0provides specialised and unique properties to\u00a0bacterium. Some of them are; \u2022 It leads to complement deposition which\u00a0provides resistance to osmotic lysis. DIAGNOSIS OF TUBERCULOSIS<\/strong>: Sputum culture<\/strong>: According to a study by Won-Jung Koh et al. a\u00a0new approach was found which can minimize the\u00a0time for tuberculosis detection on culture and drug\u00a0susceptibility testing (DST). This is known as\u00a0MGIT 960 method which decreases the turnaround\u00a0time of DST to 27 days as compared to absolute\u00a0concentration method which takes around 70 days\u00a0(Koh et al., 2012). According to a study it was\u00a0suggested that, with respect to the cost\u00a0effectiveness in India, MGIT culture is more<\/p>\n preferred over serological tests as it gives less false\u00a0positive results (Dowdy et al., 2011). Sputum\u00a0induction can be easily and safely performed in\u00a0children and infants and is more sensitive in both\u00a0HIV infected and non- infected individuals\u00a0(Hesseling et al., 2002)\u00a0An assay known as microscopic observation brothdrug\u00a0susceptibility assay (MODS) was used which\u00a0has enhanced sensitivity of around 92% as well as\u00a0uses less time for the detection of growth of\u00a0mycobacterium in liquid culture (Caviedes et al.,\u00a02000). A new advancement in this process has gain\u00a0importance. The MPB64 skin patch test\u00a0discriminates between active tuberculosis and PPD\u00a0positive healthy controls and shows very high\u00a0sensitivity of 98% with specificity 99% when\u00a0tested in Japan. In Manila, Philippines it showed\u00a0100%specificity and 88%sensitivity (Drobniweski\u00a0et al., 2003) Fluorescence-activated cell sorting<\/strong>:<\/p>\n This technique can be used in suspects with\u00a0negative AFB sputum smears. In this method BAL\u00a0cells or sputum cells are immune phenotyped. But\u00a0the frequency of region of difference-1 M.\u00a0Tuberculosis specific T cells is too low in the Polymerase Chain Reaction (PCR)<\/strong>: The problem arises when the extrapulmonary\u00a0tuberculosis is to be detected as it contains many\u00a0inhibitors which interfere in amplification. So,\u00a0proper treatment prior to reaction is necessary. So\u00a0Cobas Amplicor MTB assay was used which\u00a0eliminated all unwanted inhibitors (Bouakline et\u00a0al., 2003).<\/p>\n The PCR was used to detect mycobacteria in urine\u00a0sample using two methods- Sechi method and\u00a0Githui method. But these methods were not that\u00a0much sensitive that can be used for routine clinical\u00a0diagnosis. The sensitivity of Sechi method was\u00a028.6% and that of Githui method was 55.6%\u00a0(Kafwabulula et al., 2002).<\/p>\n The PCR was conjugated with smear microscopy to\u00a0diagnose the disease in smear-negative results. This\u00a0proved better technique by detecting two targets\u00a0devR and IS6110 sequences in the sample. These\u00a0can prove a cost effective process for rapid\u00a0diagnosis of smear-negative tuberculosis (Haldar et\u00a0al., 2007).\u00a0PCR requires proper treatment of sample prior\u00a0reaction. So a new battery operated bead beating\u00a0system is developed which can mechanically lyse\u00a0the pathogen. This device is known as OmniLyse\u00a0device developed by Claremont BioSolutions. It\u00a0takes less than 10\u00a0 minutes for the lysis of the\u00a0pathogen. It improves the safety of lysis of\u00a0mycobacteria before molecular diagnosis (Ferguson\u00a0et al., 2016).<\/p>\n Recently, a new technique of end point detection\u00a0has been used which used calcein for the visual\u00a0detection of reaction.\u00a0It binds to the manganese ions\u00a0and produces fluorescence by\u00a0 quenching effect. This made ir simpleand cost effective that can be used in resource limited developing countries Park et al<\/em>., 2003.<\/p>\n Phenotypic detection<\/strong>: Molecular Typing Techniques<\/strong>: Loop mediated isothermal amplification\u00a0(LAMP)<\/strong>: This reaction causes the production of insoluble\u00a0magnesium pyrophosphate during amplifica\u00a0which appears as turbidity (Mori and Notomi,\u00a02009).<\/p>\n The primer designing is very simple as it is done by\u00a0primer design support software\u00a0(http:\/\/primerexplorer.jp\/e\/). It automatically\u00a0designs candid primer sets specific to target\u00a0sequence. (Notomi et al., 2015).<\/p>\n The sensitivity of LAMP was increased by using\u00a0the repetitive Insertion Sequence IS6110 as a target\u00a0gene. This assay has 50 times more sensitivity then\u00a0the gyrB as a target gene (Aryan et al., 2010). The\u00a0combination of smear and LAMP is very sensitive\u00a0and can be used as in-rule test combination (George\u00a0et al., 2011).<\/p>\n LAMP has shown 100% sensitivity\u00a0for culture positive and sputum positive results\u00a0(Pandey et al., 2008).\u00a0The PURE-LAMP has very high sensitivity and\u00a0specificity which makes it economic, cost effective\u00a0and rapid method for the diagnosis of tuberculosis\u00a0(Ou et al., 2014).\u00a0LAMP can be used for the RNA as well. Reverse\u00a0transcriptase is added to the buffer and then the\u00a0reaction is carried out (RT-LAMP). (Mori and\u00a0Notomi, 2010).<\/p>\n The LAMP can be used for various applications\u00a0like genetic testing, point of care testing, rapid\u00a0testing of food products. But the major\u00a0disadvantage is the risk of production of secondary\u00a0products may be due to the high efficiency of the\u00a0reaction. So it is recommended to prevent\u00a0unwanted manipulations in master mix and reaction\u00a0tubes and detection should be done carefully and\u00a0separately. (Aryan et al., 2010).<\/p>\n In recent years, a new advancement in this\u00a0technique is use of capillary tubes to run the\u00a0reaction known as Microfluidic LAMP. The glass\u00a0capillaries are placed in between the glass slides\u00a0and the sample is injected in the capillaries. (Raftai\u00a0and Gill, 2015).<\/p>\n Mycobacterium tuberculosis resistance to\u00a0rifampicin (MTB-RIF) Assay<\/strong>:<\/p>\n In recent years, the direct nucleic acid amplification\u00a0techniques have become a potential tool for the\u00a0diagnosis of disease. In this approach, new\u00a0technique in this field is evaluated which is realtime\u00a0automated integrated system, the GeneXpert\u00a0system using the Xpert MTB\/RIF assay (GX).In\u00a0this technique the sample is firstly treated with\u00a0NaOH and isopropanol to make it inactive and then\u00a0subjected to MTB\/RIF cartridge and put in Xpert\u00a0instrument. It leads to the DNA extraction and then\u00a0amplification of 192-bp segment of the rpoB gene.\u00a0The detection is done by the hybridisation of\u00a0amplified product with the overlapping probes\u00a0which consist of core regions of rpoB gene. GX\u00a0has shown very high sensitivity for the detection of\u00a0mutations and rifampicin resistance with 100%\u00a0specificity (Moure et al., 2011).<\/p>\n The microscopic observation drug susceptibility\u00a0(MODS) is a faster method which provides Drug\u00a0Suseptibility Testing (DST). This technique\u00a0provides diagnosis as well as detects resistance\u00a0against rifampicin and isoniazid simultaneously\u00a0(Kirwan et al., 2016).<\/p>\n \u00a0In a study by Surendra Sharma et al. it was\u00a0observed that the Xpert assay detected 71% of the\u00a0\u2018\u2018confirmed TB\u2019\u2019 cases where culture and response\u00a0to anti-TB treatment were positive (Sharma et al.,\u00a02014). The pooled specificity of Xpert was 98%\u00a0(Walusimbi et al., 2013).<\/p>\n \u00a0The sensitivity for isoniazid resistance improves to\u00a090% with the advanced MTB-DRplus version of\u00a0the assay (Ling et al., 2008).<\/p>\n \u00a0The comparison was made between two real time\u00a0PCR based kits that are Cobas TaqMan MTB and\u00a0Xpert MTB\/RIF assay kit. The sensitivity of Cobas\u00a0TaqMan was less as compared to Xpert MTB\/RIF\u00a0assay kit. The Xpert MTB\/RIF was 100% sensitive\u00a0as compared to Cobas TaqMan MTB which was\u00a098% sensitive (Causse et al., 2011).<\/p>\n The high cost of Xpert and lack of impact on TB\u00a0morbidity\/mortality is a major issue (Naidoo et al.,\u00a02016). But the sensitivity and specificity of the test\u00a0make it cost effective as there is no need to do\u00a0multiple tests for diagnosis and there is reduced\u00a0number of false positive results (Vassall et al.,\u00a02011).<\/p>\n Interferon- release assay (IGRA)<\/strong>: Early Secretory Antigen Target 6 (ESAT-6) and\u00a0Culture Filtrate Protein 10 (CFP-10) are the\u00a0antigens that are highly specific for active\u00a0tuberculosis and can be used as a diagnostic reagent\u00a0(Pinxteren et al., 2000). The Quantiferon- TB Gold\u00a0test was conducted in which the whole blood\u00a0stimulated with either of the M. tuberculosis\u2013specific antigens Early Secretory Antigen Target 6\u00a0(ESAT-6) or Culture Filtrate Protein 10 (CFP-10)\u00a0was taken. The result was considered positive when\u00a0the amount of interferon- in the sample wells is\u00a0greater than or equal to 0.35 IU\/ml. It was shown\u00a0that the test has around 89% sensitivity. But it is\u00a0not yet used in routine clinical laboratories (Ferrara\u00a0et al., 2005). These tests can prove beneficial for\u00a0the diagnosis of active pulmonary tuberculosis\u00a0where there is low prevalence of latent tuberculosis\u00a0(Kang et al., 2007).<\/p>\n In a study by Nathan Shaviya et al. it was\u00a0suggested that the IFN- levels and the ratios of\u00a0IFN- to IL-10 and IFN- to adiponectin are\u00a0associated with low body weight, body mass index,\u00a0hip girth, diastolic pressure and plateletcrit\u00a0suggesting mild-inflammation during early stage\u00a0infection using newly infected TB patients.\u00a0(Shaviya et al., 2015).<\/p>\n \u00a0A new version in this field is T-SPOT assay which\u00a0is enzyme linked immunosorbent spot (ELISPOT)\u00a0assay. In this method the peripheral blood\u00a0mononuclear cells are incubated with two antigens\u00a0and then the results are detected as the number of\u00a0interferon- producing T-cells that produces spot.\u00a0If the number of spots exceeds the threshold, i.e.\u00a0usually 8, after subtracting from the number of\u00a0spots in negative control then the sample is\u00a0considered positive.<\/p>\n These tests are costly then the other tests but\u00a0economical as they generate very few false positive\u00a0results and requires only a single visit of the\u00a0individual. (Starke et al., 2014).<\/p>\n The most recent approach in this direction is the\u00a0use of electrochemical immunosensor which is\u00a0nanoparticle based. In this technique, the interferon\u00a0\u1d67 is captured by the magnetic nanoparticles and\u00a0labelled with gold nanoparticles. It is then\u00a0subjected to another anti interferon \u03b3 antibody and\u00a0multiple cadmium sulfide nanoparticles form a\u00a0complex. Then the electrochemical signals are\u00a0recorded. It takes only around 1 hour as compared\u00a0to ELISA which takes around double time. (Wang\u00a0et al., 2016) REFERENCES<\/strong>: 17. J.C. Palomino. Nonconventional and new\u00a0methods in the diagnosis of tuberculosis: feasibility\u00a0and applicability in the field. European respiratory\u00a0Journal; 26:339-350 (2005). 23. Christian Lienhardt, Knut L\u00f6nnroth, Dick\u00a0Menzies, Manica Balasegaram, Jeremiah Chakaya,\u00a0Frank Cobelens, Jennifer Cohn, Claudia M.\u00a0Denkinger, Thomas G. Evans, Gunilla K\u00e4llenius,\u00a0Gilla Kaplan, Ajay M. V. Kumar, Line\u00a0Matthiessen, Charles S. Mgone, Valerie Mizrahi,\u00a0Ya-diul Mukadi, Viet Nhung Nguyen, Anders\u00a0Nordstr\u00f6m, Christine F. Sizemore, Melvin\u00a0Spigelman, S. Bertel Squire, Soumya\u00a0Swaminathan, Paul D. Van Helden, Alimuddin\u00a0Zumla, Karin Weyer, Diana Weil, Mario\u00a0Raviglione. Translational Research for\u00a0Tuberculosis Elimination: Priorities, Challenges,\u00a0and Actions. PLoS Med; 13.3:e1001965 (2016). 40. Simon Walusimbi, Freddie Bwanga, Ayesha\u00a0DeCosta, Melles Hail,Moses Joloba and Sven Hoff\u00a0ner. Meta-analysis to compare the accuracy of\u00a0GeneXpert, MODS and the WHO 2007\u00a0algorithm for diagnosis of smear-negative\u00a0pulmonary tuberculosis. BMC infectious\u00a0diseases; 13:507 (2013). 48. Yasuyoshi Mori, Tsugunori Notomi. Loopmediated\u00a0isothermal amplification (LAMP): a\u00a0rapid, accurate and cost-effective diagnostic\u00a0method for infectious disease. Japanese Society of\u00a0Chemotherapy and the Japanese Association for\u00a0Infectious Disease; 15:62-69 (2009). \n","protected":false},"excerpt":{"rendered":" pp. 17-27 Apurva Agarwal, Gaurav Sharma and Nakuleshwar Dut Jasuja Department of Applied and Biosciences, Suresh Gyan Vihar University, Jaipur Corresponding Author: agarwalapurva00@gmail.com ABSTRACT Tuberculosis (TB) is one of the major causes of death in the world which is caused by Mycobacterium\u00a0tuberculosis. The rate of mortality is high if it is conjugated with HIV. The […]<\/p>\n","protected":false},"author":3,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[18,41],"tags":[],"class_list":["post-570","post","type-post","status-publish","format-standard","hentry","category-journal-of-environment-science-and-technology","category-volume-2-issue-1-2016-journal-of-environment-science-and-technology"],"yoast_head":"\n
\nTuberculosis is the leading cause of death in the\u00a0world which is caused by the infectious bacteria\u00a0Mycobacterium tuberculosis. It mainly affects\u00a0lungs but can affect other parts also like spine,\u00a0kidney, brain, heart. It is ranked along with HIV in\u00a0leading cause of deaths. Worldwide, 9.6 million\u00a0people are estimated to have fallen ill with TB in\u00a02014: 5.4 million men, 3.2 million women and 1.0\u00a0million children.<\/p>\n
\nMycolic acids are alpha-branched lipids that are\u00a0present in cell walls of Mycobacterium. They make\u00a0up 50% of the dry weight of the mycobacterial cell\u00a0envelope is made by the mycolic acids. Mycolic\u00a0Acids plays a significant role in virulence of\u00a0mycobacterium. They prevent attack of the\u00a0mycobacterium by cationic proteins, lysozyme, and\u00a0oxygen radicals in the phagocytic granule. Cord\u00a0factor is an inhibitor of PMN migration. It is toxic\u00a0to mammals and roduced in large amount in\u00a0virulent strains of Mycobacterium tuberculosis.\u00a0Wax-D is the major component of Freund’s\u00a0complete adjuvant (CFA).<\/p>\n
\n\u2022 High concentration leads to provide resistance\u00a0to many antibiotics.<\/p>\n
\n\u2022 Provides survival in macrophages.
\n\u2022 provides resistance against acidic and\u00a0alkaline compounds.<\/p>\n
\nThe rapid and early diagnosis of disease is required\u00a0to prevent the further infection and spread of\u00a0disease.
\nSmear microscopy<\/strong>:
\nIn most of the high burden resource poor countries,\u00a0this method is used in which acid fast bacilli are\u00a0detected in smear using a light microscope. The\u00a0smears are stained with Ziehl-Neelsen (ZN) stain.\u00a0But this technique is not so reliable. So in this\u00a0regard fluorescence microscopy can be used against\u00a0conventional light microscopy. The sensitivity was\u00a052 to 97% as compared to light microscope with\u00a0sensitivity of 32 to 94% (Parsons et al., 2011). One\u00a0of the major advantage of fluorescence microscopy\u00a0over ZN stain is that it is user friendly and the\u00a0results are unaffected by HIV status of patient\u00a0(Ndugga et al., 2003).<\/p>\n
\nIt is the most common method of diagnosis. In this\u00a0method the sputum is cultured and then the growth\u00a0of bacteria is observed. But it takes long time of\u00a0around 2-4 weeks. Sputum Induction is a new\u00a0approach which has a high yield in pulmonary\u00a0tuberculosis. Most of the patients in pleural\u00a0tuberculosis do not produce spontaneous sputum so\u00a0this method can be used for effective diagnosis\u00a0(Conde et al., 2003). Sputum induction gives best\u00a0results when used with smear microscopy and can\u00a0detect the patients who have negative spontaneous\u00a0smear microscopy. (Hepple et al., 2012).<\/p>\n
\nTuberculin skin test<\/strong>:
\nThe Tuberculosis Skin Test (TST) has been in use\u00a0for the diagnosis of tuberculosis infection since\u00a01910. TST is a protein\u2013purified derivate (PPD)\u00a0method which results from a culture filtrate of\u00a0tubercle bacilli containing over 200 antigens\u00a0common both in bacilli Chalmette-Guerin vaccine\u00a0(BCG) and in most non tuberculosis bacteria.\u00a0Therefore, the specificity and accuracy of this test\u00a0is low. Furthermore, it takes around 48-72 hours to\u00a0read TST after initial administration. It can produce\u00a0false results as error can be made in performing and\u00a0reading of results.<\/p>\n
\nDetection of Lipoarabinomannan in Urine\u00a0for Diagnosis of HIV-Associated TB<\/strong>:
\nLipoarabinomannan (LAM) is a heat-stable\u00a0glycolipid constituent in the cell wall of\u00a0Mycobacterium tuberculosis. When mycobacterium\u00a0releases this antigen it enters the circulation and\u00a0then gets filtered in renal tubes, so easily detectable\u00a0in urine of patients. It can be detected by simple\u00a0sandwich ELISA using polyclonal antibodies. It\u00a0could be used in cases where patient is infected\u00a0with HIV and it can prove a powerful tool for\u00a0diagnosis of tuberculosis in HIV patients (Achkar\u00a0et al., 2011).<\/p>\n
\nsputum to be used as stimulants for flow cytometry\u00a0cultures and other immune based assays (Lange\u00a0and Mori, 2010).<\/p>\n
\nIt was the widely used technique for the detection\u00a0of tuberculosis. It is a rapid method for the\u00a0diagnosis of disease. The technique is more\u00a0sensitive than the other previously used methods.\u00a0Out of the 144 patients that were tested it gave 80%\u00a0positive result but smear examination gave only\u00a025% accuracy (Cheng et al., 2004). In this\u00a0approach, an experiment was carried out with PCR\u00a0which was specific for devR and IS 6110\u00a0sequences. The sensitivity was 87.5% for the\u00a0combined PCR results (Chakravorty et al., 2005).<\/p>\n![\"\"](\"http:\/\/www.gyanvihar.org\/journals\/wp-content\/uploads\/2018\/12\/1-10.jpg\")
<\/p>\n![\"\"](\"http:\/\/www.gyanvihar.org\/journals\/wp-content\/uploads\/2018\/12\/1-12.jpg\")
<\/p>\n![\"\"](\"http:\/\/www.gyanvihar.org\/journals\/wp-content\/uploads\/2018\/12\/1-13.jpg\")
<\/p>\n
\nIt is a method in which lux gene is incorporated in\u00a0the phage which codes for lucifera\u00a0phage allows the detection of mycobacteriophage\u00a0by emitting light and thud allows the indirect\u00a0detection of viable Mycobacterium tuberculosis.\u00a0But this is quite expensive (Drobniewski\u00a02003).<\/p>\n
\nRestriction fragment Length Polymorphism (RFLP)\u00a0is the most commonly used technique based on\u00a0insertion sequence IS6110. It has been used to\u00a0study molecular evolution of strains, to identify\u00a0laboratory cross-contaminations, for monitoring\u00a0transmissions. (Palomino JC, 2005).<\/p>\n
\nIt is the reliable, cheap and effective technique for\u00a0the diagnosis of tuberculosis.\u00a0amplification technique which uses a
\ntemperature and a single enzyme for the detection.\u00a0It can amplify the DNA even if it is prese\u00a0small quantity. (Tomita et al.,\u00a0results in 1 hour only. It uses Bst polymerase\u00a0has polymerase activity as well as strand\u00a0displacement activity. It has two inner primers\u00a0and BIP) and two outer primers\u00a0recognise six distinct regions in the target DNA.<\/p>\n
\nIGRAs are the alternatives of tuberculin skin tests\u00a0and more sensitive and less time consuming (Santin\u00a0et al., 2012). These are blood tests which measure\u00a0ex vivo release of interferon-\u03b3 by T lymphocytes\u00a0which are stimulated by antigens specific for\u00a0Mycobacterium tuberculosis. These antigens are\u00a0not found on BCG or most non tuberculosis
\nmycobacterium so these assays are more specific\u00a0tests (Starke et al., 2014). Interferon \u03b3 release\u00a0assays are useful for children diagnosis as ii\u00a0indicates recent exposure which can later form\u00a0active tuberculosis (Rhwald et al., 2012).<\/p>\n
\nCONCLUSION<\/strong>:
\nIn recent years, the diagnostic techniques have been\u00a0advanced and became more reliable. The new\u00a0approaches are used for making diagnosis more\u00a0sensitive, economic and safe. But most of the\u00a0advanced techniques are confined to the developed\u00a0countries and are still not prevalent in developing\u00a0countries. There is a need to make them available\u00a0to the whole world so that this disease can be\u00a0diagnosed at early stage and can be declined from\u00a0the world. Tuberculosis can be eradicated by\u00a0mutual efforts, proper planning, effective diagnosis,\u00a0proper funding and continuous efforts in the field\u00a0of research and development.<\/p>\n
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