Development of a bead-based suspension array for the detection of pathogens in acute respiratory tract infections

Chen, Yu‐Sheng; Li, Hongru; Zhang, Wei; Hua, Zhi-Dan; Lin, Xiaohong; Lin, Meng-Qing; Huang, Wensen; Huang, Liping; Yu, Xinbing; Xu, Nengluan; Lin, Ming Tsan; Xie, Baosong; Shen, Xiaona; Xie, Jianfeng; Wang, Yi; Huang, Meng; Wu, Yanan; Hu, Xiang

doi:10.1177/1535370216647128

Cited by 7 publications

(8 citation statements)

References 40 publications

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“…However, there are still no commercially-available kits to detect respiratory bacterial pathogens. Recently, a Luminex assay for the detection of pathogens in acute respiratory tract infections has been built [45], however the target respiratory bacterial pathogens for this assay are limited. In addition to S. pneumoniae , S. aureus , M. pneumoniae , P. aeruginosa , and K. pneumoniae mentioned in this assay, we also involved M. catarrhalis , S. pyogenes , Legionella spp., H. influenzae as our target respiratory bacterial pathogens.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Detection of Nine Key Bacterial Respiratory Pathogens Using Luminex xTAG® Technology

Lei

Ren

Zhou

et al. 2017

IJERPH

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Early diagnosis and treatment are crucial to the outcome of lower respiratory tract infections (LRTIs). In this study, we developed an assay combining multiplex PCR and Luminex technology (MPLT) for the detection of nine important respiratory bacterial pathogens, which frequently cause LRTIs. These were Streptococcus pneumoniae, Moraxella catarrhalis, Staphylococcus aureus, Streptococcus pyogenes, Haemophilus influenzae, Mycoplasma pneumoniae, Legionella spp., Pseudomonas aeruginosa, and Klebsiella pneumoniae. Through the hybridization reaction between two new synthesized multiplex PCR products and MagPlex-TAG Microspheres, we demonstrate that the detection limits for these nine pathogens were as low as 102–103 CFU/mL. Furthermore, 86 clinical bronchoalveolar lavage fluid specimens were used to evaluate this method. Compared with the results of nine simplex real-time PCR reactions targeting these nine pathogens, this MPLT assay demonstrated a high diagnostic accuracy for Streptococcus pneumoniae (sensitivity, 87.5% and specificity, 100%). Furthermore, sensitivity and specificity for the other eight pathogens all attained 100% diagnostic accuracy. In addition, the consistency between MPLT and the nine real-time PCR reactions exceeded 98.8%. In conclusion, MPLT is a high-throughput, labor-saving and reliable method with high sensitivity and specificity for identifying nine respiratory pathogens responsible for LRTIs. Indeed, this assay may be a promising supplement to conventional methods used to diagnose LRTIs.

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Section: Discussionmentioning

confidence: 99%

Simultaneous Detection of Nine Key Bacterial Respiratory Pathogens Using Luminex xTAG® Technology

Lei

Ren

Zhou

et al. 2017

IJERPH

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“…The median fluorescence intensities (MFI) were detected using the Bio-Plex 100 suspension array system (Bio-Rad). A single multiplex PCR system was used to amplify the wzx/wzy genes as follows: an initial denaturation at 95°C for 5 min, followed by 30 cycles of denaturation at 94°C for 45 s, annealing at 50°C for 1 min, and extension at 72°C for 1min, and the final extension was done at 72°C for 10 min [ 31 ]. The PCR products were then used directly in the hybridization reaction to couple beads.…”

Section: Methodsmentioning

confidence: 99%

“…The MFIs were calculated from 100 replicate measurements using the digital signal processor and the Bio-Plex Manager software 4.1. A positive result was defined as an MFI > 150 and a signal/background (S/B = MFI/Blank) > 6.0 [ 31 ].…”

Section: Methodsmentioning

confidence: 99%

“…Such as real-time PCR assays and short sequencing assays based on serotype-specific genes [ 28 – 29 ], or chemometric analysis of attenuated total reflectance infrared spectra based on defined LPS structures [ 30 ]. Recent development of gene chip technology, including solid phase arrays and liquid bead-based suspension arrays, has given us a more sensitive and accurate method to identify bacterial serotypes [ 31 ]. The suspension array system is based on microspheres labeled with a unique dye combination.…”

Section: Introductionmentioning

confidence: 99%

“…The microspheres are coupled with specific probes for targets which are amplified from the samples using biotin-labeled primers. The fluorescent emission of the target analyte is measured by exciting the fluorescent reporter bound to the microspheres [ 31 ].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Genetic diversity of the O antigens of Proteus species and the development of a suspension array for molecular serotyping

Torzewska

Zhang

et al. 2017

PLoS ONE

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Proteus species are well-known opportunistic pathogens frequently associated with skin wound and urinary tract infections in humans and animals. O antigen diversity is important for bacteria to adapt to different hosts and environments, and has been used to identify serotypes of Proteus isolates. At present, 80 Proteus O-serotypes have been reported. Although the O antigen structures of most Proteus serotypes have been identified, the genetic features of these O antigens have not been well characterized. The O antigen gene clusters of Proteus species are located between the cpxA and secB genes. In this study, we identified 55 O antigen gene clusters of different Proteus serotypes. All clusters contain both the wzx and wzy genes and exhibit a high degree of heterogeneity. Potential functions of O antigen-related genes were proposed based on their similarity to genes in available databases. The O antigen gene clusters and structures were compared, and a number of glycosyltransferases were assigned to glycosidic linkages. In addition, an O serotype-specific suspension array was developed for detecting 31 Proteus serotypes frequently isolated from clinical specimens. To our knowledge, this is the first comprehensive report to describe the genetic features of Proteus O antigens and to develop a molecular technique to identify different Proteus serotypes.

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Early diagnosis of disease using microbead array technology: A review

Parsa

Vafajoo

Rostami

et al. 2018

Analytica Chimica Acta

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Early diagnosis of diseases (before they become advanced and incurable) is essential to reduce morbidity and mortality rates. With the advent of novel technologies in clinical laboratory diagnosis, microbead-based arrays have come to be recognized as an efficient approach, that demonstrates useful advantages over traditional assay methods for multiple disease-related biomarkers. Multiplexed microbead assays provide a robust, rapid, specific, and cost-effective approach for high-throughput and simultaneous screening of many different targets. Biomolecular binding interactions occur after applying a biological sample (such as blood plasma, saliva, cerebrospinal fluid etc.) containing the target analyte(s) to a set of microbeads with different ligand-specificities that have been coded in planar or suspension arrays. The ligand-receptor binding activity is tracked by optical signals generated by means of flow cytometry analysis in the case of suspension arrays, or by image processing devices in the case of planar arrays. In this review paper, we discuss diagnosis of cancer, neurological and infectious diseases by using optically-encoded microbead-based arrays (both multiplexed and single-analyte assays) as a reliable tool for detection and quantification of various analytes.

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Development of a bead-based suspension array for the detection of pathogens in acute respiratory tract infections

Cited by 7 publications

References 40 publications

Simultaneous Detection of Nine Key Bacterial Respiratory Pathogens Using Luminex xTAG® Technology

Simultaneous Detection of Nine Key Bacterial Respiratory Pathogens Using Luminex xTAG® Technology

Genetic diversity of the O antigens of Proteus species and the development of a suspension array for molecular serotyping

Early diagnosis of disease using microbead array technology: A review

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