Blood and urine samples as useful sources for the direct detection of tuberculosis by polymerase chain reaction

Rebollo, María José; Juan, Rafael San; Folgueira, Lola; Palenque, Elia; Díaz-Pedroche, Carmen; Lumbreras, Carlos; Aguado, José María

doi:10.1016/j.diagmicrobio.2006.03.018

Cited by 60 publications

(56 citation statements)

References 21 publications

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“…In contrast with those of other studies, (10,27) our results show that DNA amplification through PCR is a quite specific method, with reasonable sensitivity for detection of M. tuberculosis in whole blood. The performance of in-house PCR testing for TB in adult respiratory samples with negative smear microscopy test results has been described as promising, (28) although studies in detecting the existence of the disease.…”

Section: Discussioncontrasting

confidence: 53%

Desempenho da técnica nested PCR na detecção específica do complexo Mycobacterium tuberculosis em amostras sanguíneas de pacientes pediátricos

Lima

Montenegro

et al. 2009

J. bras. pneumol.

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Objective: To evaluate the performance of nested PCR (nPCR) in detecting the Mycobacterium tuberculosis complex in blood samples of patients suspected of having TB, in order to determine its potential for use as an auxiliary tool in the laboratory diagnosis of TB in children. Methods: Detection of the M. tuberculosis complex in blood samples using as a target the insertion sequence IS6110 of the genomic DNA of the bacillus. Blood samples of 120 patients were evaluated. All of the patients were under 15 years of age at the time of their treatment at public hospitals in the city of Recife, Brazil (between January of 2003 and August of 2005). Attending physicians at the hospitals diagnosed TB based on the criteria recommended by the American Thoracic Society. The nPCR amplified a 123-bp fragment with outer oligonucleotides (IS1/IS2) and, in the subsequent reaction, using inner oligonucleotides (IS3/IS4), generating an 81-bp amplicon. Results: Active or latent TB was found in 65 patients, TB was ruled out in 28 suspected cases, and 27 patients were TB-free (controls). The sensitivity of nPCR was 26.15% and was significantly higher for the extrapulmonary form of the disease (55.56%) than for the pulmonary form (18.18%). The specificity was 92.73%. Conclusions: Despite the difficulties in diagnosing TB in children and the low number of cases evaluated in the present study, nPCR in blood samples proved to be a rapid and specific technique, albeit one with low sensitivity. In order to establish its true usefulness in the diagnosis of paucibacillary forms, especially extrapulmonary TB, further studies need to be carried out with a larger sample of children and analyzing biological specimens other than blood.Keywords: Tuberculosis; Diagnosis; Blood; Polymerase chain reaction. ResumoObjetivo: Avaliar o desempenho da técnica nested PCR (nPCR) para detectar o complexo Mycobacterium tuberculosis em amostras de sangue de pacientes com suspeita de TB para sua possível utilização como uma ferramenta auxiliar no diagnóstico laboratorial da doença em crianças. Métodos: Detecção do complexo M. tuberculosis em amostras de sangue usando como alvo a sequência de inserção IS6110 do DNA genômico do bacilo. Foram avaliados 120 pacientes, menores de 15 anos de idade, de ambos os sexos, provenientes de hospitais públicos do Recife (PE), no período entre janeiro de 2003 e agosto de 2005. O diagnóstico de TB foi realizado pelo médico assistente do serviço de saúde de acordo com os critérios da Sociedade Torácica Americana. A nPCR amplificou um fragmento de 123 pb com oligonucleotídeos externos (IS1/IS2) e, na reação subsequente, com oligonucleotídeos internos (IS3/IS4), gerando um amplicon de 81 pb. Resultados: A TB ativa ou latente esteve presente em 65 pacientes, foi descartada em 28 suspeitos e 27 não tinham a doença (controles). A sensibilidade da nPCR foi de 26,15%, sendo significativamente maior na forma extrapulmonar (55,56%) em relação à pulmonar (18,18%), e a especificidade foi de 92,73%. Conclusões: Diante das dificuldades ...

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Section: Discussioncontrasting

confidence: 53%

Desempenho da técnica nested PCR na detecção específica do complexo Mycobacterium tuberculosis em amostras sanguíneas de pacientes pediátricos

Lima

Montenegro

et al. 2009

J. bras. pneumol.

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“…It is widely accepted that PCR is a new and rapid technique for the diagnosis of bacterial DNA with high specificity, even in patients who have undergone antibiotic treatment because the DNA is still detectable in the absence of a viable microorganism [7].…”

Section: Discussionmentioning

confidence: 99%

“…Several rapid methods for MTB diagnosis, such as DNA probes that require sophisticated equipment [6], have been developed. The polymerase chain reaction (PCR) is an alternative method that can amplify a small fragment of DNA with high specificity for the diagnosis of infectious diseases [7]. PCR has recently been used to detect MTB in respiratory samples [8][9][10] and other clinical specimens [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of conventional molecular diagnosis of Mycobacterium tuberculosis in clinical specimens from Morocco

Zakham

Bazoui

Akrim

et al. 2011

J Infect Dev Ctries

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Introduction: Tuberculosis is a major public health threat, annually affecting new individuals worldwide, especially those in developing countries. Rapid detection of the agent and effective treatment are two important factors in controlling this disease. Methodology: The present study aimed to evaluate polymerase chain reaction (PCR) as a rapid and direct molecular method for the diagnosis of Mycobacterium tuberculosis (MTB) in 70 clinical specimens (62 sputum samples, six cerebrospinal fluids, and two biopsies) using heat shock protein (hsp65) as the gene target. Automated sequencing of the same gene was used for the identification of MTB to the species level. Results: The sensitivity of PCR was 81.13%, with specificity of 88.24%; the positive and negative predictive values were 95.56% and 60%, respectively. Conclusion: Based on these results, the hsp65 gene sequence can be used to differentiate the members of MTB complex from nontuberculosis mycobacteria (NTM).

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“…Some of the fresh specimens that have been evaluated by various PCR include fine needle aspirates and tissue biopsies of lymph nodes, blood, urines, bone marrow aspirates and skin biopsies with variable sensitivities and specificities (Hsiao et al, 2003;Bruijnesteijn Van Coppenraet et al, 2004;Chakravorty & Tyagi, 2005;Ritis et al, 2005;Torrea et al, 2005;Rebollo et al, 2006). Although paraffin-embedded tissues are not optimal samples for PCR, often, they are the only specimens available to rule out tuberculosis.…”

Section: Laboratory-developed Pcr Methodsmentioning

confidence: 99%

Clinical Laboratory Diagnostics for Mycobacterium tuberculosis

Babady¹,

Wengenack²

2012

Understanding Tuberculosis - Global Experiences and Innovative Approaches to the Diagnosis

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Blood and urine samples as useful sources for the direct detection of tuberculosis by polymerase chain reaction

Cited by 60 publications

References 21 publications

Desempenho da técnica nested PCR na detecção específica do complexo Mycobacterium tuberculosis em amostras sanguíneas de pacientes pediátricos

Desempenho da técnica nested PCR na detecção específica do complexo Mycobacterium tuberculosis em amostras sanguíneas de pacientes pediátricos

Evaluation of conventional molecular diagnosis of Mycobacterium tuberculosis in clinical specimens from Morocco

Clinical Laboratory Diagnostics for Mycobacterium tuberculosis

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