Diagnostic

Tuberculosis Drug Resistance Detection Array Kit

The Tuberculosis Drug Resistance Detection Array Kit can detect 14 of the most frequently found mutations in three genes which are associated with resistance to the two most important (first- line) anti-tuberculosis drugs,rifampicin, and isoniazid.

Overview| Key Features| Applications| Ordering Information| Certification| Reference|

Tuberculosis (TB) remains one of the main threats to humans, causing 8 million new cases and 2 million deaths each year. The problem is becoming more critical with the emergence and spread of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis, defined as resistant to at least isoniazid (INH) and rifampin (RFP). About 2 to 3% of all new TB cases worldwide are due to MDR strain, and the highest MDR populations among new cases have been found in China (11%) and eastern Europe (7 to 14%).
China is not only one of the 22 high-burden countries that collectively accounts for ca. 80% of the world’s TB cases, but it is also the hotspot area of very high prevalence of MDR TB identified by the World Health Organization. Because of the very large financial implications of the treatment and spread of MDR strains due to globalization, MDR TB has been classified as a global pandemic more deadly than AIDS, with the potential to destabilize society.
* Drug-resistant TB emerges as a result of treatment mismanagement, and is passed from person to person in the same way as drug-sensitive TB.
* Multidrug-resistant TB (MDR-TB) is a form of TB that does not respond to the standard six month treatment using first line-drugs (i.e. resistant to isoniazid and rifampicin). It can take two years to treat with drugs that are more toxic, and 100 times more expensive. If the drugs to treat MDR-TB are mismanaged, further resistance can occur.

 

MTR Cases World Wide

% of MDR-TB among new TB cases 1994-2007


◆ Simple and versatile sample preparation: DNA extraction can be performed either from isolated strain cultures or directly from clinical sputum samples
◆ High sensitivity: our assay sensitivity is much higher than other traditional methods, because of its specially designed primer pairs and multiplex PCR procedures
◆ High accuracy: multiple data control design and optimized conditions for multiplex PCR amplification ensure the rigorous accuracy of the assay
◆ Fast analysis: the entire detection time required is less than 6 hours. This is remarkably shorter than traditional biochemical methods which require at least two weeks
◆ High consistency: strict step-by-step quality controls are used throughout the whole process, ensuring precise and reproducible results
 

Test Protocol

The DNA extracted from clinical isolates or specimens is used as a template for PCR. Because the primers are fluorescently tagged, the amplified PCR products also become fluorescently tagged. The PCR products are then hybridized to a microarray slide spotted with mutant gene test sequences, using hybridization buffer. After washing and drying, the chips are imaged using the microarray scanner. The kit interpretation software then analyzes the scan data to generate the test reports, based on the distribution of positive fluorescent probe signals on the microarray.

Recommended Work Flow
 

 

TB Drug Resitance Kit Workflow

Images of Results

TB Drug Resitance rpoB Wildtype

TB Drug Resitance rpoB T511CTB Drug Resitance rpoB C531TTB Drug Resitance rpoB C526A
rpoB WildtyperpoB T511CrpoB C531TrpoB C526A

TB Drug Resitance rpoB T533C

TB Drug Resitance katG 315AGC inhA-15CTB Drug Resitance katG 315AGC inhA-15TTB Drug Resitance katG 315ACC inhA-15C
rpoB T533CkatG 315AGC, inhA-15CkatG 315AGC, inhA-15TkatG 315ACC, inhA-15C

 

Clinical Performance

Rifampin
 

 

 
RFP DST
Sensitivity
Specificity
Coincidence
R
S
Total
Mutant
736
11
747
92.0%
97.2%
93.7%
Wild Type
64
375
439
Total
800
386
1186

Coincidence with sequencing: 100%

Isoniazid

 

 
INH DST
Sensitivity
Specificity
Coincidence
R
S
Total
Mutant
617
12
629
77.4%
96.9 %
83.8%
Wild Type
180
377
557
Total
797
389
1186

Coincidence with sequencing: 100%

Cat. No.Product NameProduct Description
301035Tuberculosis Drug Resistance Detection Array Kit12 tests

 

2

CE Certificate

Methodology

1. Guo, Y., Zhou, Y., Wang, C., Zhu, L., Wang, S., Li, Q., Jiang, G., Zhao, B., Huang, H., Yu, H., Xing W., Mitchelson, K., Cheng, J. and Zhao, Y. (2009) “Rapid and accurate determination of MDR in M. tuberculosis isolates and clinical sputum using a biochip system” International Journal of Tuberculosis and Lung Disease,13:914-920. (http://www.ncbi.nlm.nih.gov/pubmed/19555544 )
2. Chinese patent ZL200410056866.0, A asymmetrical PCR amplification method, its proprietary primers and applications.
3. Chinese patent 200810105172.X (in pending), A method to exact bacterial DNA from sputum samples and the kits and applications based on the method.