Mtp-40 and alpha antigen gene fragment amplification for the detection of Mycobacterium tuberculosis in Colombian clinical specimens

Alfonso, Rodney Alfonso; Romero, Rosa Elena; Patarroyo, Manuel Elkín; Murillo, Luis A.

doi:10.1590/s0074-02762002000800017

Cited by 5 publications

(2 citation statements)

References 30 publications

(11 reference statements)

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“…55-75%: Haas et al 2000;Zink et al 2001a;Fletcher et al 2003;Zink et al 2003). These latter data compare favourably with published rates of detection using simple PCR-based systems on modern, diagnosed, clinical samples, which are around 80% (Portillo-Gomez et al 2000;Van der Spoel van Dijk et al 2000;Mitarai et al 2001;Narayanan et al 2001;Alfonso et al 2002;Yee et al 2002;Leung et al 2003;Cheng et al 2004), and higher than detection rates for blood (40%: Taci et al 2003), urine (56%: Kafwabulula et al 2002), and host DNA in studies of animal bones from temperate archaeological sites (around 10-20%: Haynes et al 2002;Edwards et al 2004). The high frequency of amplification success from archaeological samples has been attributed to the enhanced stability of the M. tuberculosis cell wall (Donoghue et al 2004).…”

Section: Discussionsupporting

confidence: 78%

Evaluating bacterial pathogen DNA preservation in museum osteological collections

Barnes

Thomas

2005

Proc. R. Soc. B.

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Reports of bacterial pathogen DNA sequences obtained from archaeological bone specimens raise the possibility of greatly improving our understanding of the history of infectious diseases. However, the survival of pathogen DNA over long time periods is poorly characterized, and scepticism remains about the reliability of these data.In order to explore the survival of bacterial pathogen DNA in bone specimens, we analysed samples from 59 eighteenth and twentieth century individuals known to have been infected with either Mycobacterium tuberculosis or Treponema pallidum. No reproducible evidence of surviving pathogen DNA was obtained, despite the use of extraction and PCR-amplification methods determined to be highly sensitive. These data suggest that previous studies need to be interpreted with caution, and we propose that a much greater emphasis is placed on understanding how pathogen DNA survives in archaeological material, and how its presence can be properly verified and used.

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Section: Discussionsupporting

confidence: 78%

Evaluating bacterial pathogen DNA preservation in museum osteological collections

Barnes

Thomas

2005

Proc. R. Soc. B.

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“…Although the study of Sastry and Sandhya Bhat (2013) in India which was carried out on CSF specimens using IS6110 -nested PCR, 40.3% positive rate was achieved which was higher than our findings, probably due to their larger sample size, as the number of CSF specimens in this study was too few to make any conclusion. Also Alfonso et al (2002), in Colombia, reported a positivity rate of 74% by applying nested PCR targeting mtp40 on clinical specimens (urine, sputum, CSF, etc. ), while in present study 22% of the specimens were positive using mtp40 genes.…”

Section: Discussionmentioning

confidence: 99%

Identification of Mycobacterium tuberculosis in Clinical Specimens of Patients Suspected of Having Extrapulmonary Tuberculosis by Application of Nested PCR on Five Different Genes

Khosravi

Alami

Meghdadi

et al. 2017

Front. Cell. Infect. Microbiol.

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Definitive and rapid diagnosis of extrapulmonary tuberculosis (EPTB) is challenging since conventional techniques have limitations due to the paucibacillary nature of the disease. To increase the sensitivity of detection of Mycobacterium tuberculosis (MTB) in EPTB specimens, we performed a nested PCR assay targeting several genes of MTB on EPTB specimens. A total of 100 clinical specimens from suspected cases of EPTB were processed. Standard staining for acid fast bacilli (AFB) was performed as the preliminary screening test. Extracted DNAs from specimens were subjected to Nested PCR technique for the detection of five different MTB target genes of IS6110, IS1081, hsp65kd, mbp64, and mtp40. On performing AFB staining, only 13% of specimens were positive, of which ascites fluid (33.3%), followed by pleural effusion (30.8%) showed the greatest AFB positivity rate. We demonstrated slight improvement in yields in lymph node which comprised the majority of specimens in this study, by employing PCR targeted to IS6110- and hsp65-genes in comparison to AFB staining. However, the yields in ascites fluid and pleural effusion were not substantially improved by PCR, but those from bone and wound were, as in nested PCR employing either gene, the same positivity rate were obtained for ascites fluid (33.3%), while for pleural effusion specimens only IS1081 based PCR showed identical positivity rate with AFB stain (30.8%). The results for bone and wound specimens, however, demonstrated an improved yield mainly by employing IS1081 gene. Here, we report higher detection rate of EPTB in clinical specimens using five different targeted MTB genes. This nested PCR approach facilitates the comparison and the selection of the most frequently detected genes. Of course this study demonstrated the priority of IS1081 followed by mtp40 and IS6110, among the five tested genes and indicates the effectiveness of any of the three genes in the design of an efficient nested-PCR test that facilitates an early diagnosis of paucibacillary EPTB cases, which are difficult to diagnose with the available standard.

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Isolation and identification of mycobacteria in New World primates maintained in captivity

Alfonso¹,

Romero²,

Díaz³

et al. 2004

Veterinary Microbiology

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Mtp-40 and alpha antigen gene fragment amplification for the detection of Mycobacterium tuberculosis in Colombian clinical specimens

Abstract: In this study, the use of

Cited by 5 publications

References 30 publications

Evaluating bacterial pathogen DNA preservation in museum osteological collections

Evaluating bacterial pathogen DNA preservation in museum osteological collections

Identification of Mycobacterium tuberculosis in Clinical Specimens of Patients Suspected of Having Extrapulmonary Tuberculosis by Application of Nested PCR on Five Different Genes

Isolation and identification of mycobacteria in New World primates maintained in captivity

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