scholarly journals PCR-reverse blot hybridization assay in respiratory specimens for rapid detection and differentiation of mycobacteria in HIV-negative population

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Qing Zhang ◽  
Heping Xiao ◽  
Liping Yan
Keyword(s):  
Pulmonary Disease ◽  
Mixed Infection ◽  
Rapid Detection ◽  
Low Cost ◽  
Blot Hybridization ◽  
Rapid Identification ◽  
Hybridization Assay ◽  
Mycobacterial Species ◽  
High Resolution Melt ◽  
High Level

Abstract Background Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment. However, traditional method is time-consuming and cannot discriminate isolated non-tuberculosis mycobacteria (NTM) at species level. In the retrospective study, we evaluated the clinical applicability of PCR-reverse blot hybridization assay (PCR-REBA Myco-ID) with clinical specimens for rapid detection and differentiation of mycobacterial species. Methods A total of 334 sputum and 362 bronchial alveolar lavage fluids (BALF) from 696 patients with mycobacterium pulmonary disease (MPD) and 210 patients with non-mycobacterium pulmonary disease used as controls were analyzed. Sputum or BALF were obtained for MGIT 960-TBc ID test and PCR-REBA Myco-ID assay. High resolution melt analysis (HRM) was used to resolve inconsistent results of MGIT 960-TBc ID test and PCR-REBA Myco-ID assay. Results A total of 334 sputum and 362 BALF specimens from 696 MPD patients (292 MTB and 404 NTM) were eventually analyzed. In total, 292 MTBC and 436 NTM isolates (mixed infection of two species in 32 specimens) across 10 Mycobacterium species were identified. The most frequently isolated NTM species were M. intracellulare (n = 236, 54.1%), followed by M. abscessus (n = 106, 24.3%), M. kansasii (n = 46, 10.6%), M. avium (n = 36, 8.3%). Twenty-two cases had M. intracellulare and M. abscessus mixed infection and ten cases had M. avium and M. abscessus mixed infection. A high level of agreement (n = 696; 94.5%) was found between MGIT 960-TBc ID and PCR-REBA Myco-ID (k = 0.845, P = 0.000). PCR-REBA Myco-ID assay had higher AUC for both MTBC and NTM than MGIT 960-TBc ID test. Conclusion PCR-REBA Myco-ID has the advantages of rapid, comparatively easy to perform, relatively low cost and superior accuracy in mycobacterial species identification compared with MGIT 960-TBc ID. We recommend it into workflow of mycobacterial laboratories especially in source-limited countries.

scholarly journals PCR-reverse blot hybridization assay in respiratory specimens for rapid detection and differentiation of mycobacteria in HIV-negative population

2020 ◽  
Author(s):  
Qing Zhang ◽  
He-ping Xiao ◽  
Liping Yan
Keyword(s):  
Pulmonary Disease ◽  
Mixed Infection ◽  
Rapid Detection ◽  
Low Cost ◽  
Blot Hybridization ◽  
Rapid Identification ◽  
Hybridization Assay ◽  
Mycobacterial Species ◽  
Mycobacterium Species ◽  
High Level

Abstract Objective: Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment. However, traditional method is time-consuming and cannot discriminate isolated NTM to species level. In the retrospective study, we evaluated the clinical applicability of PCR-reverse blot hybridization assay (PCR-REBA) with clinical specimens for rapid detection and differentiation of mycobacterial species. Methods: A total of 334 sputum and 362 bronchial alveolar lavage fluids (BALF) from 696 patients with mycobacterium pulmonary disease and 210 patients with non-mycobacterium pulmonary disease used as controls were analyzed. Sputum samples or BALF were obtained for MGIT 960 and PCR-REBA. To confirm mycobacterium species inconsistently identified by the two different assays, high resolution melting (HRM) analysis was performed. Results: A total of 334 sputum and 362 BALF specimens from 696 patients with mycobacterium pulmonary disease (292 MTB and 404 NTM) were eventually analyzed. In total, 292 MTBC and 436 NTM isolates (co-infection of two species in 32 specimens) across 10 Mycobacterium species were identified. The most frequently isolated NTM species were M. intracellulare (n=236, 54.1%), M. abscessus (n=106, 24.3%), M. kansasii (n=46, 10.6%), M. avium (n=36, 8.3%). Twenty-two cases had mixed infection with both M. intracellulare and M. abscessus and ten cases had mixed infection with both M. avium and M. abscessus. A high level of agreement (n=696; 94.5%) was found between MGIT 960 and PCR-REBA (k = 0.845, P = 0.000). PCR-REBA had the higher AUC than MGIT 960 for both MTBC and NTM. Conclusion: PCR-REBA is helpful for rapid mycobacterial species identification with low cost and simplicity and therefore recommending its routine use.

scholarly journals A new multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay for rapid staphylococcal cassette chromosome mec (SCCmec) typing

2009 ◽  
Vol 58 (8) ◽  
pp. 1045-1057 ◽  
Author(s):  
Lin Cai ◽  
Fanrong Kong ◽  
Qinning Wang ◽  
Huiping Wang ◽  
Meng Xiao ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Blot Hybridization ◽  
Rapid Identification ◽  
Reverse Line Blot ◽  
Discriminatory Power ◽  
Hybridization Assay ◽  
Coagulase Negative Staphylococci ◽  
Primer Sets ◽  
Mrsa Strains

The aim of this study was to develop a new discriminatory method for MRSA SCCmec typing based on multiplex PCR-based reverse line-blot hybridization (mPCR/RLB) assay to enable rapid identification and classification of MRSA SCCmec types in a clinical laboratory. Forty-five primer sets and 49 probes were designed and tested in uniplex PCR (uPCR) and mPCR/RLB. Probes were compared in silico to 14 whole-genome sequences and 18 partial SCCmec gene sequences of Staphylococcus aureus and complete genome and partial SCCmec genes of seven non-MRSA strains, including meticillin-susceptible S. aureus and meticillin-resistant coagulase-negative staphylococci. The method was tested on a set of 42 well-characterized reference MRSA strains. It identified all five epidemiologically relevant SCCmec types and 26 subtypes, including established and new subtypes of SCCmec III, IV (eight subtypes each) and V (three subtypes). The discriminatory power of mPCR/RLB SCCmec typing was similar to that of MLST and spa typing (Simpson indices of diversity of 0.916, 0.926 and 0.882, respectively; differences not statistically significant). The application of mPCR/RLB hybridization assay to MRSA SCCmec typing can improve the specificity, discriminatory power and throughput of the typing procedure. The detection of up to 43 mPCR products in a single hybridization assay transforms MRSA SCCmec typing from passive epidemiological library typing into a potential tool for near-real-time infection control surveillance and tracking of MRSA transmission in hospitals.

scholarly journals Routine Use of PCR–Reverse Cross-Blot Hybridization Assay for Rapid Identification ofMycobacterium Species Growing in Liquid Media

1998 ◽  
Vol 36 (6) ◽  
pp. 1530-1533 ◽  
Author(s):  
M. Sanguinetti ◽  
B. Posteraro ◽  
F. Ardito ◽  
S. Zanetti ◽  
A. Cingolani ◽  
...  
Keyword(s):  
High Performance ◽  
Blot Hybridization ◽  
Rapid Identification ◽  
Mycolic Acid ◽  
Hybridization Assay ◽  
Clinical Samples ◽  
Liquid Media ◽  
Accurate Identification ◽  
Assay Procedure ◽  
Routine Identification

A PCR–reverse cross-blot hybridization assay procedure that is able to rapidly identify 13 species of clinically relevant mycobacteria was evaluated for routine use in the identification of acid-fast isolates growing in BACTEC 460 TB (12B and 13A) and BACTEC 9000 MB (Myco/F) liquid media. Eight of the probes used were already described by Kox et al. (L. F. F. Kox et al., J. Clin. Microbiol. 33:3225–3233, 1995). In addition, we used six other probes specific for M. chelonae, M. malmoense orM. szulgai, M. genavense,M. gordonae, M. terrae, andM. marinum/M. ulcerans that we designed ourselves. This procedure allowed us to identify 459 mycobacterial species directly from broth cultures of 5,466 clinical samples collected over 1 year and processed with the radiometric or nonradiometric BACTEC system. Our results were in agreement with those obtained by conventional identification methods and also with those obtained by mycolic acid analysis by high-performance liquid chromatography. This assay seems to be a reliable procedure for the routine identification of mycobacteria, providing an accurate identification of mycobacterial isolates more rapidly than conventional tests, with remarkable implications for an efficacious specific antimycobacterial therapy.

scholarly journals Spatial CUSUM Chart Based Method for Rapid Detection of Outbreaks

2015 ◽  
Vol 7 (1) ◽  
Author(s):  
Sesha K. Dassanayaka
Keyword(s):  
Rapid Detection ◽  
Disease Surveillance ◽  
Rapid Identification ◽  
Convincing Evidence ◽  
Cusum Chart ◽  
Simulation Studies ◽  
Disease Clusters ◽  
Spatially Correlated ◽  
False Discovery ◽  
High Level

A simple and an efficient algorithm is proposed for prospective disease surveillance using spatial CUSUSM charts. With this method, spatially correlated Poisson CUSUSM statistics are computed for small neighborhoods and the false discovery rate is controlled using the popular Benjamini-Yekutieli procedure. Simulation studies provide convincing evidence of the strength of the method in rapid identification of disease clusters. The results produced by the method are easily interpretable without a high level of statistical expertise.

Constructive Optimisation of the Propeller Type Mixing Devices Using the Virtual and Rapid Prototyping

2017 ◽  
Vol 68 (3) ◽  
pp. 453-458 ◽  
Author(s):  
Daniel Besnea ◽  
Alina Spanu ◽  
Iuliana Marlena Prodea ◽  
Gheorghita Tomescu ◽  
Iolanda Constanta Panait
Keyword(s):  
Rapid Prototyping ◽  
Low Cost ◽  
Scale Up ◽  
Three Dimensional ◽  
Rapid Identification ◽  
Multidisciplinary Optimization ◽  
External Diameter ◽  
Process Industries ◽  
Parametric Cad ◽  
Shaft Torque

The paper points out the advantages of rapid prototyping for improving the performances/constructive optimization of mixing devices used in process industries, here exemplified to propeller types ones. The multidisciplinary optimization of the propeller profile affords its design using parametric CAD methods. Starting from the mathematical curve equations proposed for the blade profile, it was determined its three-dimensional virtual model. The challenge has been focused on the variation of propeller pitch and external diameter. Three dimensional ranges were manufactured using the additive manufacturing process with Marker Boot 3D printer. The mixing performances were tested on the mixing equipment measuring the minimum rotational speed and the correspondent shaft torque for complete suspension achieved for each of the three models. The virtual and rapid prototyping method is newly proposed by the authors to obtain the basic data for scale up of the mixing systems, in the case of flexible production (of low quantities), in which both the nature and concentration of the constituents in the final product varies often. It is an efficient and low cost method for the rapid identification of the optimal mixing device configuration, which contributes to the costs reduction and to the growing of the output.

scholarly journals A comparative evaluation of two chromogenic media for surveillance of carbapenem-resistant Enterobacterales with non-carbapenemase-producing or carbapenemase-producing strains other than blaKPC: blaGES-5, blaNDM-1 or blaVIM-2

2019 ◽  
Vol 43 (3) ◽  
pp. 173-176
Author(s):  
Chang-Hun Park
Keyword(s):  
Rapid Detection ◽  
Comparative Evaluation ◽  
Susceptibility Testing ◽  
Rapid Identification ◽  
Surveillance Program ◽  
Contact Precaution ◽  
Chain Reaction ◽  
Carbapenem Resistant ◽  
Control And Prevention ◽  
Polymerase Chain

Abstract Background Infections caused by carbapenem-resistant Enterobacterales (CREs) are an emerging problem associated with high rates of morbidity and mortality. CREs are divided into two categories (carbapenemase-producing [CP] CRE and non-CP CRE). The most prevalent carbapenemase produced by Enterobacterales is Klebsiella pneumoniae carbapenemase (KPC) in Korea. Rapid identification of CREs is clinically important in infection control precaution. We compared the performance of two chromogenic media (chromID CARBA agar and CHROMagar KPC agar) for non-CP CREs or CP CREs with blaGES-5, blaNDM-1 or blaVIM-2 in a Korean hospital. Methods The study was carried out during a 3-month period from April to June 2017 during the surveillance program for CRE colonization. Antimicrobial susceptibility testing (AST) and polymerase chain reaction (PCR) were performed at the Korean Centers for Disease Control and Prevention. Results A total of 45 rectal swabs from 42 hospitalized patients were examined. Sensitivity of both chromID CARBA and CHROMagar KPC were 100% for CP CREs; and 50% and 100% for non-CP CREs, respectively. Specificity of chromID CARBA and CHROMagar KPC were 89.2% and 70.3% for CP CRE, respectively; and 76.9% and 66.7% for non-CP CRE, respectively. Conclusions The CHROMagar KPC is useful to monitor non-CP and CP CREs. The chromID CARBA is efficient for rapid detection of CP CREs requiring high contact precaution.

scholarly journals Hybrid mosquitoes? Evidence from rural Tanzania on how local communities conceptualize and respond to modified mosquitoes as a tool for malaria control

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Marceline F. Finda ◽  
Fredros O. Okumu ◽  
Elihaika Minja ◽  
Rukiyah Njalambaha ◽  
Winfrida Mponzi ◽  
...  
Keyword(s):  
Malaria Control ◽  
Technology Development ◽  
Low Cost ◽  
Public Support ◽  
Community Leaders ◽  
Community Perceptions ◽  
Novel Technologies ◽  
Control Programmes ◽  
High Level ◽  
Interpretive Frames

Abstract Background Different forms of mosquito modifications are being considered as potential high-impact and low-cost tools for future malaria control in Africa. Although still under evaluation, the eventual success of these technologies will require high-level public acceptance. Understanding prevailing community perceptions of mosquito modification is, therefore, crucial for effective design and implementation of these interventions. This study investigated community perceptions regarding genetically-modified mosquitoes (GMMs) and their potential for malaria control in Tanzanian villages where no research or campaign for such technologies has yet been undertaken. Methods A mixed-methods design was used, involving: (i) focus group discussions (FGD) with community leaders to get insights on how they frame and would respond to GMMs, and (ii) structured questionnaires administered to 490 community members to assess awareness, perceptions and support for GMMs for malaria control. Descriptive statistics were used to summarize the findings and thematic content analysis was used to identify key concepts and interpret the findings. Results Nearly all survey respondents were unaware of mosquito modification technologies for malaria control (94.3%), and reported no knowledge of their specific characteristics (97.3%). However, community leaders participating in FGDs offered a set of distinctive interpretive frames to conceptualize interventions relying on GMMs for malaria control. The participants commonly referenced their experiences of cross-breeding for selecting preferred traits in domestic plants and animals. Preferred GMMs attributes included the expected reductions in insecticide use and human labour. Population suppression approaches, requiring as few releases as possible, were favoured. Common concerns included whether the GMMs would look or behave differently than wild mosquitoes, and how the technology would be integrated into current malaria control policies. The participants emphasised the importance and the challenge of educating and engaging communities during the technology development. Conclusions Understanding how communities perceive and interpret novel technologies is crucial to the design and effective implementation of new vector control programmes. This study offers vital clues on how communities with no prior experience of modified mosquitoes might conceptualize or respond to such technologies when deployed in the context of malaria control programmes. Drawing upon existing interpretive frames and locally-resonant analogies when deploying such technologies may provide a basis for more durable public support in the future.

scholarly journals Sunflower-Like Nanostructure with Built-In Hotspots for Alpha-Fetoprotein Detection

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1197
Author(s):  
Xiaoyu Zhao ◽  
Aonan Zhu ◽  
Yaxin Wang ◽  
Yongjun Zhang ◽  
Xiaolong Zhang
Keyword(s):  
Rapid Detection ◽  
Clinical Medicine ◽  
Electromagnetic Coupling ◽  
Low Cost ◽  
Surface Enhanced Raman Spectroscopy ◽  
Alpha Fetoprotein ◽  
Low Concentrations ◽  
Surface Enhanced Raman ◽  
Active Substrate ◽  
Array Structures

In the present study, a sunflower-like nanostructure array composed of a series of synaptic nanoparticles and nanospheres was manufactured through an efficient and low-cost colloidal lithography technique. The primary electromagnetic field contribution generated by the synaptic nanoparticles of the surface array structures was also determined by a finite-difference time-domain software to simulate the hotspots. This structure exhibited high repeatability and excellent sensitivity; hence, it was used as a surface-enhanced Raman spectroscopy (SERS) active substrate to achieve a rapid detection of ultra-low concentrations of Alpha-fetoprotein (AFP). This study demonstrates the design of a plasmonic structure with strong electromagnetic coupling, which can be used for the rapid detection of AFP concentration in clinical medicine.

Sign in / Sign up

Export Citation Format

Share Document