Detection of Mycobacterium Tuberculosis DNA in Blood of Patients with Acute Pulmonary Tuberculosis by Polymerase Chain Reaction and Non-Isotopic Hybridisation Assay

Prete, Raffaele Del; D'Alagni, Marina; Sabato, Roberto; Picca, Vito; Miragliotta, G.

doi:10.1099/00222615-46-6-495

Cited by 16 publications

(15 citation statements)

References 26 publications

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“…Blood testing has also been explored by FOLGUEIRA et al [15] who showed a sensitivity of 82% in HIV-infected and of 33% in HIV-seronegative patients with an 8-fold greater sensitivity than M. tuberculosis cultures. Similar results were obtained by DEL PRETE et al [16] who amplified the IS6110 sequence in 86% of 30 blood samples from TB patients and by GAMBOA et al [17] who amplified M. tuberculosis RNA from 90% of TB patient blood samples with the amplified mycobacteria direct test (AMTD).…”

Section: Saltinisupporting

confidence: 71%

“…It tells us that more work must be done to adapt amplification tests to each type of specimen that needs to be examined in order to diagnose extrapulmonary TB. In the context of the reports on M. tuberculosis DNA amplification from blood samples quoted above [14][15][16][17], the fact that the IS6110 test used by KOLK et al [18] correctly diagnosed almost all of the pus specimens and a good percentage of the pleural fluids, but virtually none of the blood samples suggests that specimen preparation, in addition to DNA amplification can still be improved.…”

Section: Saltinimentioning

confidence: 99%

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Direct amplification of Mycobacterium tuberculosis deoxyribonucleic acid in paucibacillary tuberculosis

Saltini¹

1998

Eur Respir J

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The discovery of the polymerase chain reaction (PCR) as a means to generate detectable signals from as little as a single molecule of target deoxyribonucleic acid (DNA) [1] has led to the development of extremely powerful and rapid diagnostic tests for a number of infectious agents. PCR was therefore expected to improve significantly the sensitivity of tests for the diagnosis of tuberculosis (TB), compared with the standard stains for acid fast bacilli (AFB) on sputum smears. Since the first report by HANCE et al.[2] on the use of PCR amplification of the 65 kDa heat shock protein (HSP) gene for the diagnosis of Mycobacterium tuberculosis in clinical samples, a number of M. tuberculosis DNA sequences have been tested as amp-lification targets [3]. Over the past ten years, several PCR test formats have been employed and nested PCR tests have been described which were endowed with a detec-tion power 1,000-fold greater (lower detection limit, 0.1 M. tuberculosis colony-forming unit (cfu)·mL -1 ) than that of the standard PCR with 35 amplification cycles (detection limit, 100 cfu·mL -1 ) [4].Among DNA targets, the IS6110 insertion sequence, a M. tuberculosis complex-specific sequence which is repeated 10-16 times in the chromosome of M. tuberculosis [5], has been the most widely used. This test has the power of amplifying a fraction of a chromosome of M. tuberculosis and with this test it has been possible to accomplish something as spectacular as identifying mycobacteria buried in mummies thousands of years old [6]. However, the test is difficult to handle and its reproducibility across laboratories has been rather disappointing [7]. Even more importantly, all the amplification tests using either the PCR or other ribonucleic acid (RNA) and DNA amplification techniques that have been developed, bottled and marketed for the diagnosis of M. tuberculosis in routine laboratories do not seem sufficiently sensitive for the diagnosis of paucibacillary TB. Notwithstanding their extreme power in the laboratory setting and although the rapid amplification tests are more sensitive than AFB stains, home grown and commercial tests alike, both still fail to diagnose about 50% of microscopy-negative sputum samples [8] i.e., those samples containing less than the 5-10,000 M. tuberculosis organisms·mL -1 of sample that are required for a positive AFB smear.With this background, an American Thoracic Society (ATS) panel recently recommended that the rapid amplification tests should be used for the confirmation of rapid diagnosis of M. tuberculosis versus nontuberculous mycobacteria in individuals with an AFB-positive sputum smear. With a direct amplification test it can be rapidly determined (with >95% sensitivity) whether an individual has active, multibacillary TB (requiring isolation and contact screening), or a nontuberculous mycobacterial infection (not requiring any of the above). In contrast, the same ATS panel could not recommend the use of rapid amplification tests for the routine diagnosis of paucibacillary TB, due to thei...

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Section: Saltinisupporting

confidence: 71%

Section: Saltinimentioning

confidence: 99%

Direct amplification of Mycobacterium tuberculosis deoxyribonucleic acid in paucibacillary tuberculosis

Saltini¹

1998

Eur Respir J

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“…A PCR na detecção de micobactérias não é à prova de falhas, pois sua sensibilidade na detecção de micobactérias tem sido relatada na faixa de 55-100% [28][29][30][31] . Por outro lado, esta investigação pode ser usada quando a carga bacteriana é muito baixa 15,32 .…”

Section: Discussionunclassified

Children with significant cervical lymphadenopathy: clinicopathological analysis and role of fine-needle aspiration in Indian setup

Khan¹,

Wahab²,

Chana³

et al. 2008

J Pediatr (Rio J)

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ResumoObjetivo: Estudar o perfil clínico-patológico de crianças indianas com linfadenopatia cervical e o papel da citologia aspirativa por agulha fina com ênfase especial na tuberculose como causa. Resultados: A hiperplasia reativa foi o tipo mais comum de linfadenite, seguida da granulomatosa. Os linfonodos do triângulo posterior unilateral foram o grupo afetado com maior freqüência no grupo de linfadenopatia cervical tuberculosa. A aspiração por agulha fina, seguida da coloração de Ziehl-Neelsen, histopatologia e cultura em associação, obteve sucesso em realizar o diagnóstico em 85,7% dos casos de etiologia tuberculosa. Métodos Conclusões:A aspiração por agulha fina é uma ferramenta diagnóstica valiosa no tratamento de crianças com apresentação clínica de linfonodos cervicais aumentados. A técnica reduz a necessidade de procedimentos mais invasivos e dispendiosos, principalmente em países em desenvolvimento.Cultura e histopatologia, entretanto, devem ser consideradas em casos nos quais a citologia aspirativa por agulha fina não é diagnóstica.J Pediatr (Rio J). 2008;84(5):449-454: Linfadenopatia cervical, citologia aspirativa por agulha fina, crianças. AbstractObjective: To study the clinicopathological profile of children from India with cervical lymphadenopathy and the role of fine-needle aspiration cytology with special emphasis on tuberculosis as a cause. Methods:A total of 89 children in the age group of 10 months to 12 years, presenting to our hospital from April 2004 to March 2005, were included. All the patients underwent thorough clinical and investigational assessment vis-à-vis cervical lymphadenopathy. Outcome measurements included clinical status and ability of conventional tests to categorize different types of lymphadenopathy and their utility in diagnosing tubercular lymphadenitis. Interobserver variability was analyzed measuring kappa test and was found to be in agreement. Results:Reactive hyperplasia was the most common type of lymphadenitis, followed by granulomatous involvement. Unilateral posterior triangle lymph nodes were the most commonly affected in the tubercular cervical lymphadenopathy group. Fine-needle aspiration followed by Ziehl-Neelsen staining, histopathology and culture in combination were able to perform the diagnosis in 85.7% of cases affected with tubercular etiology. Conclusions:Fine-needle aspiration is a valuable diagnostic tool in the management of children with the clinical presentation of enlarged cervical lymph nodes. The technique reduces the need for more invasive and costly procedures, especially in a Third World country. Culture and histopathology, however, should be considered in cases where repeated fine-needle aspiration cytology is non-diagnostic.J Pediatr (Rio J). 2008;84(5):449-454: Cervical lymphadenopathy, fine-needle aspiration cytology, children.

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“…Este tipo de pruebas están basadas en la presencia del microorganismo en sangre (bacteriemia) o, quizá más importante, la presencia de fragmentos de ADN de la micobacteria que está parcialmente digerida por macrófagos que la han fagocitado en el sitio de la infección y, luego, han entrado a la circulación (41)(42)(43). Este evento parece ser común en pacientes con sida e infectados con M. tuberculosis o M. avium, pero también se ha reportado en pacientes sin un inmunocompromiso importante (41)(42)(43)(44)(45)(46)(47)(48). La detección de ADN de la micobacteria en sangre periférica, podría complementar aquellas estrategias que dependen de la respuesta inmune del paciente.…”

Section: ¿Cuál Es El Método De Elección?unclassified

Nuevas herramientas para la detección de la tuberculosis latente.

Restrepo¹

2004

biomedica

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La tuberculosis es una causa importante de muerte en el mundo, principalmente en países en vía de desarrollo. Se estima que cada año hay 9 millones de casos nuevos y 1,5-3 millones de muertes (1,2). Esta infección es causada por Mycobacterium tuberculosis, una bacteria ácido- La tuberculosis es un problema serio de salud pública en el mundo, especialmente en países en vía de desarrollo, inclusive Colombia. En nuestro país estamos enfocados en el manejo clínico de los pacientes con tuberculosis activa, pero no hay campañas efectivas que identifiquen y provean terapia a los individuos con las formas latentes de la infección y con alto riesgo de progresar hacia la enfermedad. En esta revisión se plantea la importancia de realizar dichas campañas para evitar la diseminación de la infección en la comunidad. Esto incluye la búsqueda activa y el tratamiento profiláctico de los contactos de casos recientes, así como la de individuos con tuberculosis latente con alto riesgo de desarrollar la enfermedad. En ausencia de una prueba de oro para detectar la tuberculosis latente, la prueba cutánea de la tuberculina se ha utilizado por más de 100 años para dicho fin, a pesar de sus limitaciones de sensibilidad y especificidad. En esta revisión se evalúan las ventajas y desventajas de una nueva generación de inmunoensayos que incluye la prueba comercial Quantiferon y el desarrollo experimental del ELISPOT. Ambas se basan en la detección de IFNγ secretado por linfocitos de sangre periférica cuando se incuban con antígenos específicos del bacilo tuberculoso. Finalmente, se plantea la importancia de desarrollar pruebas moleculares enfocadas en detectar el ADN de la micobacteria como posible complemento a los inmunoensayos descritos.

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Detection of Mycobacterium Tuberculosis DNA in Blood of Patients with Acute Pulmonary Tuberculosis by Polymerase Chain Reaction and Non-Isotopic Hybridisation Assay

Cited by 16 publications

References 26 publications

Direct amplification of Mycobacterium tuberculosis deoxyribonucleic acid in paucibacillary tuberculosis

Direct amplification of Mycobacterium tuberculosis deoxyribonucleic acid in paucibacillary tuberculosis

Children with significant cervical lymphadenopathy: clinicopathological analysis and role of fine-needle aspiration in Indian setup

Nuevas herramientas para la detección de la tuberculosis latente.

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