Multichannel Series Piezoelectric Quartz Crystal Research Articles

Electrochemical biosensor for rapid detection of bacteria based on facile synthesis of silver wire across electrodes

The early detection of bacteria is of critical importance in addressing serious public health problems. Here, an electrochemical biosensor for rapid detection of bacteria based on facile synthesis of silver wire across electrodes was constructed. High-variable region of 16S rRNA of bacteria was used as biomarker. Polymerase-free synthesis of silver wire was introduced into electrochemical signal transduction to improve the sensitivity of electrochemical detection. The construction biosensor of proposed method is as follows: Metastable hairpin probe H1 was modified on electrode surface, biomarker can open the stem-loop structure of H1 and activates HCR. The alternate opening of the stem-loop structure of H1 and H2-AuNPs finally results in the formation of long double-stranded DNA-RNA (HCR products) -AuNPs. The formed AuNPs modified HCR products was blown in one direction using N2 to across the electrode gap. Using this HCR products as template, the silver wire was formed between the electrodes by silver deposition, and resulted in sharp change in electrical parameters of electrode. As the proof-of-concept work, multichannel series piezoelectric quartz crystal (MSPQC) was utilized as detector. The detection of Staphylococcus aureus in the concentration range from 50 to 107 CFU/mL within 100 min was achieved. The detection limit was 50 CFU/mL. Escherichia coli, Salmonella enteritidis, Listeria innocua, Pseudomonas aeruginosa and Streptococcus pneumoniae did not interfere the detection results. This newly proposed electrochemical biosensor is simple, rapid and exhibit high signal-to-noise ratio, it has great potential for being applied in food safety monitoring and clinical diagnosis.

The construction of Mycobacterium tuberculosis 16S rDNA MSPQC sensor based on Exonuclease III-assisted cyclic signal amplification

Tuberculosis caused by Mycobacterium tuberculosis (M. tuberculosis) remains one of the most serious infectious diseases all over the world. The key to reduce the spread and mortality rate of tuberculosis is to develop faster and more sensitive approaches for detection of M. tuberculosis. However, current detection methods can not meet the requirements of rapid clinical M. tuberculosis detection in terms of detection time. Herein, a new 16S rDNA multichannel series piezoelectric quartz crystal (MSPQC) sensor based on Exonuclease III (Exo III)-aided target recycling has been developed for rapid detection of M. tuberculosis. The specific 16S rDNA fragment of M. tuberculosis was used as biomarker, DNA capture probes complementary to the biomarker were designed and modified on the surface of AuNPs. The Exo III which could recognise hybrid duplexes and selectively digest DNA capture probe was used to assist digestion cycle by digesting DNA capture probe and releasing the intact target fragment. After all DNA probes loading on the surface of AuNPs were removed, the surface of AuNPs was exposed and conductive connection was formed between the nanogap network electrode by self-catalytic growth of exposed AuNPs in the glucose and HAuCl4 solution. This resulted in sensitive response of M. tuberculosis sensor and M. tuberculosis was detected by recording this response. The limit of detection (LOD) of the method was 20 CFU/mL and the detection time was less than 3 h. It was expected to be widely used in detection methods of M. tuberculosis.

A supersensitive MSPQC bacterium sensor based on 16S rRNA and “DNA-RNA switch”

The early detection of bacterium plays a significant role in addressing serious public health issues. In this paper, a supersensitive multichannel series piezoelectric quartz crystal (MSPQC) sensor of bacterium based on 16S rRNA and “DNA-RNA switch” was constructed. The fragment in specific region of 16S rRNA was used as the biomarker of bacterium to ensure high specificity and to achieve the accurate judgment of microbial vitality. “DNA-RNA switch” was designed to conduct two electrodes by switching insulated “gene-link” into conductive “silver-link”, which achieved the super-sensitivity of MSPQC to bacteria. To demonstrate the feasibility of this strategy, a proof-of-concept method for Escherichia coli (E. coli) assay was designed. The detection limit was down to 2 cfu/mL. Staphylococcus aureus, Salmonella enteritidis, Listeria innocua and Pseudomonas aeruginosa did not interfere with the detection results. Proposed method was highly sensitive, and specific for bacterium detection, which might find widely use in early detection of bacterium in the field of public safety monitoring and clinical diagnosis.

Mycobacterium tuberculosis strain H37Rv Electrochemical Sensor Mediated by Aptamer and AuNPs-DNA.

The accurate and rapid detection of Mycobacterium tuberculosis ( M. tuberculosis) is essential for the effective treatment of tuberculosis. In this article, we propose an electrochemical sensor to detect M. tuberculosis reference strain H37Rv. The sensor contains an H37Rv aptamer and oligonucleotides modified with gold nanoparticles (AuNPs-DNA). An H37Rv aptamer screened by our laboratory was used as the recognition probe. The change in frequency shift mediated by AuNPs-DNA in the presence of H37Rv was detected using a multichannel series piezoelectric quartz crystal (MSPQC) system. Three oligonucleotides modified with gold nanoparticles were designed. These oligonucleotides contained 12, 12, and 13 bases that hybridized with the 37-nt H37Rv aptamer. H37Rv aptamer was immobilized on the gold electrode surface by Au-S bonds. A conductive-layer was then formed by sequential hybridization of the aptamer with the three designed AuNPs-DNAs. When H37Rv was present, it specifically bound to the aptamer, resulting in the detachment of AuNPs-DNA from the electrode. The conductive layer was thereby replaced by a nonconductive complex of aptamer and bacteria. These changes were monitored by the MSPQC system. The proposed sensor is rapid, specific and sensitive, the detection time was 2 h. The detection limit was 100 cfu/mL. This sensor would be of great benefit for the early clinical diagnosis of tuberculosis.

A new aptamer/polyadenylated DNA interdigitated gold electrode piezoelectric sensor for rapid detection of Pseudomonas aeruginosa

Rapid detection of Pseudomonas aeruginosa (P. aeruginosa) is of great importance for accurate diagnosis and treatment of infected patients. In this study, a novel method was developed for the selective detection of P. aeruginosa by combing the sandwich type complex of magnetic bead/aptamer/polyadenylated-DNA with the sensitive detection platform of gold (Au) interdigital electrode connected to a multichannel series piezoelectric quartz crystal (Au IDE-MSPQC) system. Here, the magnetic bead (MB) was used as carrier for immobilization of the aptamer of P. aeruginosa. Polyadenylated DNA was bound to the aptamer through complementary strand pairing. When the P. aeruginosa was present in the sample solution, the polyadenylated DNA was replaced by the P. aeruginosa because of the specific interaction between P. aeruginosa and its aptamer. The released polyadenylated DNA strand in the detected solution could adsorb onto the surface of Au IDE by virtue of the strong interaction between adenine (A) and Au IDE, and result in sensitive frequency shift response of the MSPQC sensor. The limits of detection (LOD) of the method were as low as 9 CFU/mL in buffer and 52 CFU/mL in simulated blood sample. The proposed method was successfully applied to the selective detection of P. aeruginosa in blood samples. The constructed sensor is expected to find application for the rapid detection of P. aeruginosa in environment, food and clinical diagnosis.

Selection of a new Mycobacterium tuberculosis H37Rv aptamer and its application in the construction of a SWCNT/aptamer/Au-IDE MSPQC H37Rv sensor

A rapid and accurate detection method for Mycobacterium tuberculosis (M. tuberculosis) is essential for effectively treating tuberculosis. However, current detection methods cannot meet these clinical requirements because the methods are slow or of low specificity. Consequently, a new highly specific ssDNA aptamer against M. tuberculosis reference strain H37Rv was selected by using the whole-cell systematic evolution of ligands by exponential enrichment technique. The selected aptamer was used to construct a fast and highly specific H37Rv sensor. The probe was produced by immobilizing thiol-modified aptamer on an Au interdigital electrode (Au-IDE) of a multichannel series piezoelectric quartz crystal (MSPQC) through Au-S bonding, and then single-walled carbon nanotubes (SWCNTs) were bonded on the aptamer by π-π stacking. SWCNTs were used as a signal indicator because of their considerable difference in conductivity compared with H37Rv. When H37Rv is present, it replaces the SWCNTs because it binds to the aptamer much more strongly than SWCNTs do. The replacement of SWCNTs by H37Rv resulted in a large change in the electrical properties, and this change was detected by the MSPQC. The proposed sensor is highly selective and can distinguish H37Rv from Mycobacterium smegmatis (M. smegmatis) and Bacillus Calmette-Guerin vaccine (BCG). The detection time was 70min and the detection limit was 100cfu/mL. Compared with conventional methods, this new SWCNT/aptamer/Au-IDE MSPQC H37Rv sensor was specific, rapid, and sensitive, and it holds great potential for the early detection of H37Rv in clinical diagnosis.

Novel S-MSPQC cell sensor for real time monitoring the injury of endothelial cell by LPS and assessing the drug effect on this injury

A novel square Au microelectrode multi-channel series piezoelectric quartz crystal (S-MSPQC) cell sensor was constructed by square Au microelectrode in series connected with quartz crystal sensor in MSPQC. The experimental results showed square shape Au microelectrode was more sensitive than normally used interdigital microelectrode. New constructed S-MSPQC was successfully used for real time monitoring the injury of human umbilical vein endothelial cells which induced by lipopolysaccharide (LPS) and assessing the drug effects of two anti-oxidative vitamins, VC and VE and their combination against this injury. The detection results showed that the low concentration of LPS may promote the proliferation of the HUVEC cells at first 12h, then it caused the cell apoptosis. The pretreatment of VC or VE on the cells for 12h before adding the LPS could partially reduced the injury of LPS on HUVEC cells. The sensor detection results were all consisted with the results detected by MTT assay and microscopy observation. The synergistic effect of the combination of VC-VE on LPS-induced cell injury was also observed by sensor. Compared with the traditional biological methods, the proposed method is sensitive, label-free, non-invasive, cheap, simple, and automatic. It provides a new automatic tool for cell monitor in the field of cell biology, cytobiology, and drug effects research.

Multichannel series piezoelectric quartz crystal cell sensor for real time and quantitative monitoring of the living cell and assessment of cytotoxicity.

A new multichannel series piezoelectric quartz crystal (MSPQC) cell sensor for real time monitoring of living cells in vitro was reported in this paper. The constructed sensor was used successfully to monitor adhesion, spreading, proliferation, and apoptosis of MG63 osteosarcoma cells and investigate the effects of different concentrations of cobalt chloride on MG63 cells. Quantitative real time and dynamic cell analyses data were conducted using the MSPQC cell sensor. Compared with methods such as fluorescence staining and morphology observation by microscopy, the MSPQC cell sensor is noninvasive, label free, simple, cheap, and capable of online monitoring. It can automatically record the growth status of cells and quantitatively evaluate cell proliferation and the apoptotic response to drugs. It will be a valuable detection and analysis tool for the acquisition of cellular level information and is anticipated to have application in the field of cell biology research or cytotoxicity testing in the future.

A New Bacteriophage-Modified Piezoelectric Sensor for Rapid and Specific detection of Mycobacterium

Detection of tuberculosis and related diseases caused by mycobacteria is costly, time-consuming, and labor-intensive. Here a new phage-modified piezoelectric system for rapid and specific detection of mycobacteria was developed. In this system, interdigital gold electrode immobilized with lytic phage was used as a probe in place of a steel electrode in the multi-channel series piezoelectric quartz crystal (MSPQC) system. The probe was directly connected to the piezoelectric detection system. Mycobacterium was specifically captured to the phage-modified electrode and then lysed by immobilized phage, which caused the electrode electrical properties change. This change can be sensitively monitored by the piezoelectric detection system. The detection time of Mycobacterium smegmatis was less than 2 hours and a detection limit of 103cfu mL−1 was obtained. Additionally, it was successfully used to detect Mycobacterium tuberculosis. The developed system using phage-modified interdigital electrode showed high specificity and reproducibility for mycobacterium detection. Compared with the MSPQC system, the proposed system was faster and more specific.

Multi-Channel Series Piezoelectric Quartz Crystal Sensor with Partial Least Square Method for Identification of Clinically Relevant Bacteria

Multi-Channel Series Piezoelectric Quartz Crystal Sensor with Partial Least Square Method for Identification of Clinically Relevant Bacteria

A New Sensor Method for Studying the Effect of Gentamicin and Cefotaxime Combination Against Escherichia coli

A new sensor method based on a multi-channel series piezoelectric quartz crystal (MSPQC) was proposed for studying the effect of gentamicin and cefotaxime combination against Escherichia coli ATCC 25922. The frequency curves under different combination regimens were obtained and compared with each other. When antibiotic had an inhibitory effect to the growth of bacteria, frequency detection time (FDT) would be prolonged. The FDT was used to assess the effect of an antibiotic combination. By using the proposed method, simultaneous administration, different order, and time interval of nonsimultaneous administration were investigated in detail. The MSPQC method can provide the process information in real time. It is simple, rapid, and easy to perform.

New MSPQC Method for Rapid Identification and Quantification of Pseudomonas aeruginosa

A new method for the rapid identification and quantification of Pseudomonas aeruginosa using multichannel series piezoelectric quartz crystal (MSPQC) was proposed. The identification of P. aeruginosa was based on the development of acetamide broth, which can selective culture P. aeruginosa and performed perfectly in MSPQC. The quantitative detection of P. aeruginosa was based on the fact that the frequency detection time (FDT) detected by MSPQC in developed medium had a linear relationship with the logarithm of its initial concentration in the range of 10–108 colony − forming units (cfu) ml–1 (R=−0.984). The detection limit was 10 cfu ml–1.

A new MSPQC for rapid growth and detection of Mycobacterium tuberculosis

Many of the deaths caused by tuberculosis (TB) in the world are due to wrong or late diagnosis. The World Health Organization (WHO) calls for better and cheaper TB tests method for this reason. In this paper, a new multi-channel series piezoelectric quartz crystal (MSPQC) sensor system was developed for rapid growth and detection of Mycobacterium tuberculosis. It is an automatic continuous monitoring system. The system was used to detect TB based on the volatile metabolic products NH 3 and CO 2 during the growth of M. tuberculosis. The metabolic products, diffusing from the medium into the KOH absorbing solution, resulted in the conductance change of the absorbing solution detected by the MSPQC sensitively. The frequency shift versus time response curves were recorded by self-developed software. Frequency detection time (FDT) corresponding to −100 Hz in frequency shift value was used as a parameter to quantitatively determine M. tuberculosis H37Ra (an avirulent strain). H37Ra and 40 strains clinic positive samples were detected by the proposed system successfully. As for H37Ra, the FDT had a linear relationship with the logarithm of its initial concentration in the range of 10 2–10 7 colony forming units (cfu) ml −1 ( R = −0.998) and the detection limit was low to 10 cfu ml −1. 4% NaOH solution that can kill contaminating microorganisms and make M. tuberculosis alive was used as pretreatment reagent to provide selectivity to this method. Comparative tests were also carried out by using BACTEC™ MGIT™ 960 and conventional Lowenstein–Jensen (L–J) slants. The results showed that the proposed system was quicker than BACTEC™ MGIT™ 960 and it is also cheaper and will be widely used in TB tests in the world.

A new B-PAn-P system for the detection of bacteria population

A new base-type polyaniline piezoelectric bacteria culture (B-PAn-P) system for rapid detection of bacteria population was proposed. In this system, interdigital electrode coated with a base-type polyaniline/poly vinyl alcohol (B-PAn/PVA) composite film was used as probe (B-PAn probe) in place of steel electrode in multi-channel series piezoelectric quartz crystal (MSPQC) system. The probe was directly connected to the piezoelectric detection system. B-PAn/PVA film can react with CO2 produced by the growth of bacteria in culture medium, which caused the film electrical properties change. This change can be sensitively monitored by piezoelectric detection system. The superiority of using proposed probe instead of steel electrode were: firstly, it was more sensitive to bacteria growth response and secondly, it did not require a specially formulated culture medium which was demanded in MSPQC system. Just like MSPQC, proposed system can be used to detect bacteria population. When used to quantitatively determine Pseudomonas aeruginosa, the frequency detection time (FDT) obtained in frequency shift curve had a linear relationship with the logarithm values of initial concentration of P. aeruginosa in the range of 10 to 106cfuml−1. The detection limit was 10cfuml−1. Compared with MSPQC system, proposed system was more sensitive and no special demand to culture medium.

A new antimicrobial susceptibility testing method of Escherichia coli against ampicillin by MSPQC

A new antimicrobial susceptibility testing method by multi-channel series piezoelectric quartz crystal (MSPQC) was proposed. This method was used to test susceptibility of clinical Escherichia coli isolates against ampicillin. Both the minimum inhibitory concentrations (MICs) and interpretive categorization of clinical E. coli isolates were determined by proposed method. Comparing tests were run at the same time by the agar dilution method and the disk diffusion method. The experimental results showed that MSPQC method had a good agreement with the reference methods. Compared with those methods, the MSPQC method is simple, rapid, and convenient to perform. It can offer both a minimum inhibitory concentration (MIC) and an interpretive category result.

A new MSPQC system for rapid detection of pathogens in clinical samples

A new multi-channel series piezoelectric quartz crystal (MSPQC) system for detection of pathogens in clinical sample was proposed. Some factors, which affect the detection of pathogens by using MSPQC, were all investigated. A total of 650 clinical samples were detected by MSPQC and compared with licensed BACTEC 9120 system (Becton Dickinson Diagnostic Instrument Systems, Sparks, MD, USA) simultaneously in the Third Xiangya Hospital of Central South University, China. When the incubation period was 5 days, two systems had similar detected results: the MSPQC system detected 123 growth of 650 (18.92%) bottles while the BACTEC 9120 detected 125 growth of 650 (19.23%) bottles. The MSPQC had 2 false-positive signals and 2 false-negative signals. However, BACTEC 9120 had 3 false-positive signals and 0 false-negative signals. Further identifications of bacteria were run by VITEK-2 (bioMérieux China Ltd.), 5% sheep blood trypticase soy agar (SBA) and chocolate agar (CA). Comparing with BACTEC 9120, MSPQC system possesses following advantages: shorter average detection time, less blood volume needed, less false-positive results and low cost. It can also provide information in real time. So MSPQC has a wonderful perspective in clinical application.

An improving method for determination of Escherichia coli population based on multi-channel series piezoelectric quartz crystal system

A new application of conductometry based on multi-channel series piezoelectric quartz crystal (MSPQC) is described for quantification of the bacteria population and the antibiotic effect. The result of MSPQC system showed a good linear relation ( r=−0.9885) between the frequency detection time (FDT) and its initial concentration of Escherichia coli, and the antibiotic that effected on the metabolism activity of E. coli had a significant variation of slope as a quantification index.