The Capilia TB-Neo assay (an immunochromatographic test) was performed to all repeat-culture positive samples in accordance with the manufacturer’s instructions (TAUNS Laboratories, Inc., Shizuoka, Japan) to differentiate MTB complex from NTM infections. The Capilia TB-Neo detects MPT64 protein secreted by species of MTB complex. Isolates of MTB complex detected by Capilia TB-Neo were further subjected to a GenoType MTBC assay (Hain Lifescience, Nehren, Germany) for confirmation and identification to the species level. In order to identify species that were Capilia TB-Neo negative (NTM suspects), 2 additional genotyping methods: the GenoType CM/AS (Hain Lifescience) were performed according to the manufacturer’s instructions. GenoType CM can simultaneously detect and differentiate up to 27 clinically relevant NTM species from MTBC. GenoType AS can detect and differentiate an additional 19 NTM species.
Genotype mtbc assay
Manufactured by Hain Lifescience
Sourced in Germany
The GenoType MTBC assay is a molecular genetic test used for the identification of the Mycobacterium tuberculosis complex (MTBC) from clinical specimens. The assay utilizes DNA strip technology to detect the presence of MTBC DNA, without providing further species-level differentiation within the MTBC.
Automatically generated - may contain errors
Lab products found in correlation
Genotype cm as, by Hain Lifescience (1 mentions)
Bactec mgit 960 system, by BD (1 mentions)
Accuprobe, by Hologic (1 mentions)
Oxalic acid, by Merck Group (1 mentions)
Stonebrink, by BD (1 mentions)
L wenstein jensen, by BD (1 mentions)
Minimix, by Interscience (1 mentions)
Genolyse isolation kit, by Hain Lifescience (1 mentions)
5 protocols using genotype mtbc assay
1
Rapid Identification of Mycobacterial Infections
2
M. tuberculosis Complex Identification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All isolates were identified as members of the M. tuberculosis complex, using gene probes (ACCUProbe; Hologic, San Diego, CA, USA) or the GenoType MTBC assay (Hain Lifescience, Nehren, Germany). The differentiation between the species of the M. tuberculosis complex was based on genotyping data of each individual isolate [7 (link)–9 (link)]. Drug susceptibility testing was determined by means of the proportioning method on Löwenstein–Jensen medium and/or by means of the modified proportioning method with the BACTEC MGIT 960 system (Becton Dickinson Microbiology Systems, Cockeysville, MD, USA).
3
Identification of M. tuberculosis Complex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After culture growth, the AccuProbe M. tuberculosis complex culture identification test (Gen-Probe Inc., San Diego, California) was performed according to the manufacturer's instructions and was used as the “gold standard” for the identification of M. tuberculosis complex species in the primary isolation. Because these samples were obtained from a country that has the highest prevalence of Mycobacterium africanum recorded in the African continent [22 (link)] and the AccuProbe M. tuberculosis complex assay is not able to discriminate the species within the M. tuberculosis complex, the Genotype MTBC assay (Hain Lifescience, Nehren, Germany) was applied for the differentiation within the M. tuberculosis complex from cultured material as recommended by the manufacturer.
4
Mycobacterium Culture and Genotyping Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The culture was performed according to the standard procedure used at the National Veterinary Research Institute (NVRI) in Pulawy (Poland). Briefly, the collected material was decontaminated with a 5% solution of oxalic acid (Sigma-Aldrich, Saint Louis, MO, USA). Homogenization was performed in a stomacher (MiniMix, Interscience, France) for three minutes (1500 g, 10 min). The supernatant was removed and the pellet was washed twice with sterile physiological sodium chloride solution (Polfa, Lublin, Poland).
The obtained pellet was seeded on two solid media: Stonebrink (Becton Dickinson, Franklin Lakes, NJ, USA) and Löwenstein–Jensen (Becton Dickinson, Franklin Lakes, NJ, USA). The media were incubated at 37 °C. Growth was assessed every seven days for 12 weeks.
The DNA was isolated from the cultures and the genotype determined as described previously [1 (link)]. Briefly, DNA isolation was performed using a Genolyse isolation kit (Hain Lifescience, Nehren, Germany). Strains were determined using the GenoType®MTBCassay (Hain Lifescience, Nehren, Germany).
The obtained pellet was seeded on two solid media: Stonebrink (Becton Dickinson, Franklin Lakes, NJ, USA) and Löwenstein–Jensen (Becton Dickinson, Franklin Lakes, NJ, USA). The media were incubated at 37 °C. Growth was assessed every seven days for 12 weeks.
The DNA was isolated from the cultures and the genotype determined as described previously [1 (link)]. Briefly, DNA isolation was performed using a Genolyse isolation kit (Hain Lifescience, Nehren, Germany). Strains were determined using the GenoType®MTBCassay (Hain Lifescience, Nehren, Germany).
5
Profiling TB Granulomas from Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole tissue sections were obtained from 14 human lung TB granulomas (Supplementary Table 1 for details). Sample selection was based on histological evidence of TB and PCR positivity for Mtb DNA. All TB granulomas samples were collected from HIV-negative patients. FFPE tissue samples were obtained with informed patients' consent and retrieved from the pathology files of the Clínica Universidad de Navarra (protocol 111/2010). Histological evidence of TB granulomas with central necrosis surrounded by a highly cellular area at the periphery were evaluated based on the review of hematoxylin and eosin-stained sections from FFPE blocks by two authors of this study (CEA and MAM). In addition, the detection of acid-fast bacilli (AFB) was investigated in all cases by the conventional Ziehl-Neelsen method, as described elsewhere [10] . The presence of Mtb DNA in tissue samples was performed as part of the routine diagnostic guidelines of the Clínica Universidad de Navarra by using a commercially available multiplex PCR-based, solid-phase reverse hybridization GenoType MTBC assay (Hain Lifescience GmbH, Nehren, Germany) [11] .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!