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Evaluation of FLASH - PCR forrapid detection of Mycobacterium tuberculosis from clinical specimens.

Imani Fooladi AA, Tarvedi Zadeh Y, Mehrab R, Halabian R, Azizi T - Iran J Microbiol (2013)

Bottom Line: The findings of this study have suggested that removal of the contaminants in FLASH-PCR sign ificantly reduced the detection time, and MTB was much more rapidly detected in the clinical specimens compared to the conventional culture and smear examination.Results of the medical survey showed that the majority of TB patients were males, over 51 years old, smokers, with pulmonary TB and normal chest X-ray (CXR).MTB can be rapidly detected inclinical specimens using FLASH-PCR in comparison with culture and smear examination.

View Article: PubMed Central - PubMed

Affiliation: Applied Microbiology, Research Center Baqiyatallah University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background and objectives: Tuberculosis (TB) is the oldest known bacterial disease in humans. Due to the rise of morbidity in recent years, early diagnosis of the disease is necessary.

Materials and methods: In this study we used Fluorescent Amplification-Based Specific Hybridization (FLASH) PCR to targetIS6110 for rapid detection of M. tuberculosis (MTB). To investigate the important factors influencing the risk of TB, data from patients and their medical records were analyzed.

Result: The sensitivity and specificity of FLASH-PCR for detecting MTB were determined as 93.33% and 92.5%, respectively. The findings of this study have suggested that removal of the contaminants in FLASH-PCR sign ificantly reduced the detection time, and MTB was much more rapidly detected in the clinical specimens compared to the conventional culture and smear examination. Results of the medical survey showed that the majority of TB patients were males, over 51 years old, smokers, with pulmonary TB and normal chest X-ray (CXR).

Conclusion: MTB can be rapidly detected inclinical specimens using FLASH-PCR in comparison with culture and smear examination.

No MeSH data available.


Related in: MedlinePlus

FLASH-PCR; upper fig: chart of flash PCR that detected with GeneXpert software.Lower fig: gel electrophoresis; M: 100bp ladder, 2 &7 were negative sample, 1 was low positive sample.
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Figure 1: FLASH-PCR; upper fig: chart of flash PCR that detected with GeneXpert software.Lower fig: gel electrophoresis; M: 100bp ladder, 2 &7 were negative sample, 1 was low positive sample.

Mentions: FLASH-PCR was performed with IS6110 as specific target gene for MTB detection. For detection of FLASH-PCR products, PCR product tubes were transferred to Fluorescent Detector FD-12. The PCR reaction was validated by detection of fluorescent IC. In positive samples (MTB patients) fluorescence of MTB probe was detected (Fig. 1). Therefore, the products are easily identified without the extra step of electrophoresis and the risk of contamination.


Evaluation of FLASH - PCR forrapid detection of Mycobacterium tuberculosis from clinical specimens.

Imani Fooladi AA, Tarvedi Zadeh Y, Mehrab R, Halabian R, Azizi T - Iran J Microbiol (2013)

FLASH-PCR; upper fig: chart of flash PCR that detected with GeneXpert software.Lower fig: gel electrophoresis; M: 100bp ladder, 2 &7 were negative sample, 1 was low positive sample.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4385165&req=5

Figure 1: FLASH-PCR; upper fig: chart of flash PCR that detected with GeneXpert software.Lower fig: gel electrophoresis; M: 100bp ladder, 2 &7 were negative sample, 1 was low positive sample.
Mentions: FLASH-PCR was performed with IS6110 as specific target gene for MTB detection. For detection of FLASH-PCR products, PCR product tubes were transferred to Fluorescent Detector FD-12. The PCR reaction was validated by detection of fluorescent IC. In positive samples (MTB patients) fluorescence of MTB probe was detected (Fig. 1). Therefore, the products are easily identified without the extra step of electrophoresis and the risk of contamination.

Bottom Line: The findings of this study have suggested that removal of the contaminants in FLASH-PCR sign ificantly reduced the detection time, and MTB was much more rapidly detected in the clinical specimens compared to the conventional culture and smear examination.Results of the medical survey showed that the majority of TB patients were males, over 51 years old, smokers, with pulmonary TB and normal chest X-ray (CXR).MTB can be rapidly detected inclinical specimens using FLASH-PCR in comparison with culture and smear examination.

View Article: PubMed Central - PubMed

Affiliation: Applied Microbiology, Research Center Baqiyatallah University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background and objectives: Tuberculosis (TB) is the oldest known bacterial disease in humans. Due to the rise of morbidity in recent years, early diagnosis of the disease is necessary.

Materials and methods: In this study we used Fluorescent Amplification-Based Specific Hybridization (FLASH) PCR to targetIS6110 for rapid detection of M. tuberculosis (MTB). To investigate the important factors influencing the risk of TB, data from patients and their medical records were analyzed.

Result: The sensitivity and specificity of FLASH-PCR for detecting MTB were determined as 93.33% and 92.5%, respectively. The findings of this study have suggested that removal of the contaminants in FLASH-PCR sign ificantly reduced the detection time, and MTB was much more rapidly detected in the clinical specimens compared to the conventional culture and smear examination. Results of the medical survey showed that the majority of TB patients were males, over 51 years old, smokers, with pulmonary TB and normal chest X-ray (CXR).

Conclusion: MTB can be rapidly detected inclinical specimens using FLASH-PCR in comparison with culture and smear examination.

No MeSH data available.


Related in: MedlinePlus