Download RQ-MBT Complex Technical leaflet

REALQUALITY
REALQUALITY
RQ-MBT Complex
Compatible platforms: Validated on
! Applied Biosystems 7500 Fast / Fast Dx, 7300 and
StepOnePlus / StepOne Real-Time PCR System
!
LightCycler® 480 and 2.0 Real-Time PCR System
(Roche)
!
Dx Real-Time System and CFX96 Real-Time PCR
Detection System (Bio-Rad)
code RQ-85
Kit for detection and quantification of the
Mycobacterium tuberculosis complex
(MTBC) by Real-Time PCR
INTRODUCTION
Mycobacteria are gram positive, non-motile, acid-fast, nonspore forming and obligate aerobe or microaerophilic. The
genus Mycobacterium comprises the Mycobacterium
tuberculosis complex (MTBC) and more than 100 other
species of ubiquitous nontuberculous mycobacteria (atypic
and NTM) which include pathogenic as well as nonpathogenic and opportunistic species.
According to the World Health Organization report of 2007
(WHO report, 2007), also nowadays tuberculosis (TB)
occurs worldwide and poses an enormous problem for the
public health.
Although the pulmonary tuberculosis is the most common
type, basically all organs can be affected. About 73 % of TB
cases suffer from pulmonary tuberculosis. In 3.5 % of those
cases other organs are affected as well.
Although, the initial diagnosis of an infection usually is
based on clinical data, the definitive diagnosis requires
isolation and identification of the mycobacterium in the
laboratory. The conventional methods for mycobacterium
detection are based on analysis of phenotype, biochemical
characteristics and on chromatography and are usually too
time-consuming to be applied in routine screening. In the
last years, several methods for direct detection of the
mycobacteria have been developed that are based on
techniques of molecular biology. Among these, the
methods based on PCR allow detection of the
mycobacterium at a fraction of time and cost. Due to the
fact that such PCR-based tests do not provide information
about the vitality of the pathogen and may give positive
results also after culture negativization or completion of the
therapy, they are to be used exclusively in the diagnostic
phase and are not suited for therapy follow-up.
TECHNICAL CHARACTERISTICS
The kit can be used on instruments that allow a reaction
volume of 25 µL and read the FAM and JOE fluorescence.
Analytical specificity: No non-specific pairing of primers
and probes, no cross-reaction
Analytical sensitivity (detection limit): 0.5 copies of
bacterial genome / µL of DNA extract
7
Analytical sensitivity (linear range): 5 – 10 copies of
bacterial genome / reaction
Reproducibility, intra-assay variability: 0.468 %
Reproducibility, inter-assay variability: 0.417 %
Diagnostic specificity: 97 %
Diagnostic sensitivity: 96 %
Accuracy: 96 %
Fig. 1: MTBC standard curve, analyzed on the Applied Biosystems
7500 Real-Time PCR System using SDS software.
ORDERING INFORMATION
Code
Product
PKG
RQ-85-48/96
REALQUALITY
RQ-MBT Complex
48/96 tests
For creating standard curves:
Code
Product
PKG
RQ-86-ST
REALQUALITY
RQ-MBT Complex
STANDARD
4 × 60 µL
Number of tests: 48 or 96
Shelf life: 12 months
Sample material: DNA extract from sputum, BAL, FFPE
tissue and cultures in liquid growth medium
Amplified region: gene region coding for the 16S
ribosomal RNA
Internal Control: Amplification of β-globin gene in
multiplex with target pathogen
Positive controls: DNA fragment from the MTBC genome
and the β-globin gene
REFERENCES
Tortoli E. et al., Micobatteriologia Clinica :11, 2008.
Ministero della salute. Epidemiologia della tubercolosi in Italia anni 1995-2005, 2007.
Stauffer F, Haber H, Rieger A, et al. J Clin Microbiol 36: 614-617,
1998.
Moore D.F., Curry J.I., Knott C.A., Jonas V, J.Clin.Microbiol.
34:1745-1749, 1996.
RQ-MBT_Complex_techleaf_e20140718.doc
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