Rapid diagnosis of tuberculous meningitis: A comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, MPB-64 and 65 kDa antigen

Rafi, Wasiulla; Venkataswamy, Manjunatha M.; Ravi, Vasanthapuram; Chandramuki, A

doi:10.1016/j.jns.2006.11.001

Cited by 40 publications

(39 citation statements)

References 32 publications

(35 reference statements)

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“…Some studies have assessed amplification of multiple gene targets from the same samples, but this was using CSF [15]; an increase in sensitivity was noted when all these assays were analysed together [16]. A study by Hulder et al [17] reported an increase in sensitivity of PCR if two targets were used for diagnosis of TBM.…”

Section: Discussionmentioning

confidence: 99%

Does multiplex Polymerase Chain Reaction increase the diagnostic percentage in osteoarticular tuberculosis? A prospective evaluation of 80 cases

Sharma

et al. 2011

International Orthopaedics (SICOT)

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Purpose Multiplex Polymerase Chain Reaction (MPCR) is a technique in which two or more gene targets are amplified in a single reaction. This has increased sensitivity of diagnosis as a single gene target may be absent in some Mycobacterium tuberculosis strains. Methods MPCR using two target genes specific for Mycobacterium tuberculosis, that is, IS6110 and MPB 64, ZN staining and Mycobacterial culture were performed on synovial fluid/pus samples of 80 (three confirmed, 77 suspected) patients of osteoarticular tuberculosis and 25 non tuberculosis patients. Results MPCR had a sensitivity of 100% in confirmed cases and 81.8% in clinically suspected cases. AFB was positive in one patient and Mycobacterial culture was positive in three patients. MPCR also had 100% specificity; MPB64 was positive in five patients in which IS6110 was negative whereas IS6110 was positive in two patients in which MPB64 was negative.Conclusions MPCR is a sensitive and specific method for diagnosis of paucibacilliary conditions such as osteoarticular tuberculosis.

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Section: Discussionmentioning

confidence: 99%

Does multiplex Polymerase Chain Reaction increase the diagnostic percentage in osteoarticular tuberculosis? A prospective evaluation of 80 cases

Sharma

et al. 2011

International Orthopaedics (SICOT)

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“…Experimentally, PCR assays are able to detect M. tuberculosis DNA down to an estimated 2-CFU/ml concentration (10). Many of these assays utilize primers directed at the insertion sequence IS6110 (30,77,102,137,166,167,180,240), which is a repetitive element exclusively found in the genome of the M. tuberculosis complex (230), although isolates with low copy numbers and even no copy numbers have been identified (230). A recent example of this method includes the use of an IS6110 uniplex PCR assay that had 98% sensitivity (NPV, 99%) and a specificity of 100% (PPV ϭ 100%) against the "gold standard" of culture (167) and an overall sensitivity of 76.4% (NPV, 59.9%) and specificity of 89.2% (PPV, 94.7%) when clinical TBM was included (166).…”

Section: Molecular and Biochemical Analysismentioning

confidence: 99%

“…Many of these assays utilize primers directed at the insertion sequence IS6110 (30,77,102,137,166,167,180,240), which is a repetitive element exclusively found in the genome of the M. tuberculosis complex (230), although isolates with low copy numbers and even no copy numbers have been identified (230). A recent example of this method includes the use of an IS6110 uniplex PCR assay that had 98% sensitivity (NPV, 99%) and a specificity of 100% (PPV ϭ 100%) against the "gold standard" of culture (167) and an overall sensitivity of 76.4% (NPV, 59.9%) and specificity of 89.2% (PPV, 94.7%) when clinical TBM was included (166). Several methods of PCR have been evaluated, including quantitative nested real-time PCR (208)(209)(210), the Amplicor Mycobacterium tuberculosis Test (23,24), and a variety of other in-house PCR assays (45,46,51,83,104,111,120,123,138,141,165,189,193).…”

Section: Molecular and Biochemical Analysismentioning

confidence: 99%

Central Nervous System Tuberculosis: Pathogenesis and Clinical Aspects

et al. 2008

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SUMMARY Tuberculosis of the central nervous system (CNS) is a highly devastating form of tuberculosis, which, even in the setting of appropriate antitubercular therapy, leads to unacceptable levels of morbidity and mortality. Despite the development of promising molecular diagnostic techniques, diagnosis of CNS tuberculosis relies largely on microbiological methods that are insensitive, and as such, CNS tuberculosis remains a formidable diagnostic challenge. Insights into the basic neuropathogenesis of Mycobacterium tuberculosis and the development of an appropriate animal model are desperately needed. The optimal regimen and length of treatment are largely unknown, and with the rising incidence of multidrug-resistant strains of M. tuberculosis, the development of well-tolerated and effective antibiotics remains a continued need. While the most widely used vaccine in the world largely targets this manifestation of tuberculosis, the BCG vaccine has not fulfilled the promise of eliminating CNS tuberculosis. We put forth this review to highlight the current understanding of the neuropathogenesis of M. tuberculosis, to discuss certain epidemiological, clinical, diagnostic, and therapeutic aspects of CNS tuberculosis, and also to underscore the many unmet needs in this important field.

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“…Sin embargo, esta apreciación no es exacta y la sensibilidad y especifi cidad de esta técnica es muy variable y depende de factores como una óptima extracción del ADN, si se realiza el procedimiento "en casa" ("in house") o con un kit comercial y la correcta aplicación de los pasos para la reacción de polimerasa en cadena (RPC); todo ello relacionado con la experiencia del operador Existen diversas secuencias de ADN específi cas de M tuberculosis que pueden ser amplifi cadas como la IS986, Ag85, MPB-64, TRC4, 65 kDa y 38 kDa 31 . Sin embargo, la IS6110 ha demostrado ser el ensayo de RPC de un solo paso de mejor sensibilidad y especifi cidad [32][33][34] .…”

Section: Biología Molecular En El Diagnóstico De Mtbcunclassified

Meningitis tuberculosa: claves para su diagnóstico y propuestas terapéuticas

2011

Rev. chil. infectol.

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IntroducciónP ara el año 2006 se estimaron en 9,2 millones los nuevos casos de tuberculosis (TBC) en el mundo con un incremento de su incidencia de 0,4% por año 1 . La meningitis tuberculosa (MTBC) representa el 1% anual de todos los casos de TBC 2 y su mortalidad es elevada, llegando a ser de 30% en aquellos pacientes que reciben tratamiento óptimo 3 . El más importante factor que infl uencia en el pronóstico de la MTBC es el diagnóstico precoz y el inicio temprano de tratamiento específi co 4 .La mayoría de las veces en las que un médico clínico se ve enfrentado a un paciente con una probable MTBC, establecer un diagnóstico certero y buscar una terapia realmente efi ciente para cada caso representa un desafío.Considerada la forma más letal de TBC 5 , en la práctica su diagnóstico se establece por una suma de elementos clínicos y de laboratorio que la mayoría de veces son inespecífi cos, los cultivos son de crecimiento tardío y buena parte de los casos son tratadas sin un sustento microbiológico estableciéndose un "diagnóstico terapéutico" cuando existe una buena respuesta a la terapia anti TBC, lo que representa una moneda al aire para el médico tratante.En este contexto se presenta esta revisión de las principales herramientas disponibles para el diagnóstico y las más razonables opciones de tratamiento de la MTBC; además se realizan propuestas prácticas para el manejo de estos pacientes. Diagnóstico de meningitis tuberculosa Manifestaciones clínicasLa presentación de la MTBC suele ser sub-aguda y habitualmente toma más de 7 días, siendo en las zonas de alta incidencia propia de individuos jóvenes [6][7][8] .Los síntomas y signos principales, basados en las series clínicas de MTBC más grandes 7-9 , se señalan en la Tabla 1. Destaca la fi ebre, cefalea, compromiso de conciencia, signos de irritación meníngea e hiponatremia. Se advierte lo inhabitual de las crisis convulsivas con sólo dos eventos descritos en la serie de Roca de un total de 29 casos de MTBC en un período de 15 años 9 . La gravedad de la MTBC se clasifi ca dentro de tres estados, de acuerdo al método de Gordon y Parson 10 los que se hallan relacionados con la duración de los síntomas previos, mortalidad y secuelas 7 (Tabla 2).Cerca de 75% de los pacientes con MTBC tienen concomitantemente TBC en otra localización 11,12 y ~60% puede llegar a tener hallazgos radiológicos consistentes con TBC pulmonar 7 .Meningitis tuberculosa: claves para su diagnóstico y propuestas terapéuticas Martín Lasso B.Tuberculous meningitis: tips for diagnosis and proposals for treatment Diagnosis of tuberculous meningitis (TBM) is always a challenge. We must give importance for duration of clinical manifestations. Cerebrospinal fl uid (CSF) has own characteristic and it have to be control several times during the treatment. Adenosin deaminase with cut off more than 15 UI/mL and M. tuberculosis polymerase chain reaction in CSF are the most relevant diagnostic tests. Radiologic test gives diagnostic clues but do not confi rm the diagnosis. In the future we can structure a s...

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Rapid diagnosis of tuberculous meningitis: A comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, MPB-64 and 65 kDa antigen

Cited by 40 publications

References 32 publications

Does multiplex Polymerase Chain Reaction increase the diagnostic percentage in osteoarticular tuberculosis? A prospective evaluation of 80 cases

Does multiplex Polymerase Chain Reaction increase the diagnostic percentage in osteoarticular tuberculosis? A prospective evaluation of 80 cases

Central Nervous System Tuberculosis: Pathogenesis and Clinical Aspects

Meningitis tuberculosa: claves para su diagnóstico y propuestas terapéuticas

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