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1.
Virol J ; 21(1): 171, 2024 Aug 01.
Article in English | MEDLINE | ID: mdl-39090721

ABSTRACT

BACKGROUND: This study aimed to demonstrate that the genomic material of SARS-CoV-2 can be isolated from strips of COVID-19 rapid diagnostic test cassettes. METHOD: It was a prospective cross-sectional study involving patients admitted to treatment centers and sampling sites in the city of Conakry, Guinea. A total of 121 patients were double sampled, and 9 more patients were tested only for RDT. PCR was conducted according to the protocol of the RunMei kit. Sequencing was performed by using the illumina COVIDSeq protocol. Nine COVID-19 RDTs without nasopharyngeal swabs were in addition tested. RESULT: Among the 130 COVID-19 RDTs, forty-seven were macroscopically positive, whereas seventy-two were positive according to PCR using RDT strip, while among the 121 VTM swabs, sixty-four were positive. Among eighty-three negative COVID-19 RDTs, twenty-seven were positive by PCR using RDT strip with a geometric mean Ct value of 32.49 cycles. Compared to those of PCR using VTM, the sensitivity and specificity of PCR using RDT strip were estimated to be 100% and 85.96%, respectively, with 93.39% test accuracy. Among the fifteen COVID-19 RDT extracts eligible for sequencing, eleven had sequences identical to those obtained via the standard method, with coverage between 75 and 99.6%. CONCLUSION: These results show that COVID-19 RDTs can be used as biological material for the genomic surveillance of SARS-CoV-2.


Subject(s)
COVID-19 Nucleic Acid Testing , COVID-19 , RNA, Viral , SARS-CoV-2 , Adult , Female , Humans , Male , Middle Aged , COVID-19/diagnosis , COVID-19/virology , COVID-19 Nucleic Acid Testing/methods , Cross-Sectional Studies , Diagnostic Tests, Routine/methods , Genome, Viral/genetics , Nasopharynx/virology , Prospective Studies , Rapid Diagnostic Tests/instrumentation , Reagent Strips , RNA, Viral/genetics , RNA, Viral/isolation & purification , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Sensitivity and Specificity
2.
Molecules ; 29(13)2024 Jun 27.
Article in English | MEDLINE | ID: mdl-38998998

ABSTRACT

Cancer is one of the major public health challenges in the world, which is characterized by rapid progression and high mortality. Immunotherapy, represented by PD-1 monoclonal antibody, has significantly improved the efficacy of malignant tumors and has become one of the most popular immunotherapy methods at present. Therefore, there is an increasing demand for novel detection methods for PD-1 monoclonal antibodies. The aim of this work was to establish a rapid, simple, and sensitive immunochromatographic test strip (ICTS) based on the AuNPs enlargement for both visual and instrumental detection of the PD-1 monoclonal antibody concentration. The mixed solution of NH2OH·HCl and HAuCl4 was used as an enhancement solution to lower the detection limit and achieve higher sensitivity. A test strip reader was used to construct a visualized quantitative detection standard curve for the PD-1 monoclonal antibody concentration. The LOD was 1.58 ng/mL through a triple signal-to-noise ratio. The detection time was within 10 min. The constructed test strips can rapidly, accurately, and efficiently detect the concentration of PD-1 monoclonal antibody in real samples.


Subject(s)
Antibodies, Monoclonal , Chromatography, Affinity , Metal Nanoparticles , Programmed Cell Death 1 Receptor , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/chemistry , Programmed Cell Death 1 Receptor/immunology , Chromatography, Affinity/methods , Metal Nanoparticles/chemistry , Humans , Gold/chemistry , Reagent Strips , Limit of Detection
3.
BMJ Open ; 14(7): e081830, 2024 Jul 16.
Article in English | MEDLINE | ID: mdl-39013655

ABSTRACT

OBJECTIVE: The utilisation of pH level measurements from gastric contents may indicate the preferred tip position of a nasogastric tube or monitor the efficacy of stress ulcer prophylaxis in critically ill patients. We aimed to determine the accuracy of pH strip (pHS) tests and pH liquid (pHL) tests compared with the standard pH meter (pHM). DESIGN: Diagnostic accuracy study. SETTING: Gastric contents from medically critically ill patients. PARTICIPANTS: In total, 113 gastric samples were collected from 27 critically ill patients. OUTCOME MEASURE: The level of pH measured by pHM, pHS and pHL. RESULTS: The pH values measured by pHM, pHS and pHL were 5.83 (IQR 5.12-6.61), 5.50 (IQR 5.00-6.00) and 5.75 (IQR 5.25-6.25), respectively. The pHS test showed greater accuracy, exhibiting a more positive correlation with the standard pHM measurement than the pHL test, with Y=0.95*X+0.56; rho=0.91, p<0.001, and Y=1.09*X - 0.72; rho=0.75, p<0.001, respectively. However, the pHS test demonstrated less agreement with the pHM than the pHL test, with biases of -0.27 versus 0.18, respectively. Noticeably, a slight variation in pHL from the standard pH values was found when we measured gastric contents with a pH lower than 5. CONCLUSION: Both the pHS and pHL methods were good options for measuring gastric pH in critically ill patients. However, it was advisable to find alternative approaches to the pHL testing method when anticipated gastric acidity levels fall below 5. TRIAL REGISTRATION NUMBER: TCTR20220530004.


Subject(s)
Critical Illness , Gastric Acidity Determination , Gastrointestinal Contents , Humans , Female , Male , Hydrogen-Ion Concentration , Middle Aged , Gastrointestinal Contents/chemistry , Aged , Reagent Strips , Adult , Reproducibility of Results
4.
Harm Reduct J ; 21(1): 115, 2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38877522

ABSTRACT

BACKGROUND: Fentanyl test strips (FTS) are lateral flow immunoassay strips designed for detection of ng/mL levels of fentanyl in urine. In 2021, the US Centers for Disease Control and the Substance Abuse and Mental Health Administration stated that federal funds could be used for procurement of FTS for harm reduction strategies approved by the government such as drug checking. The market for FTS has expanded rapidly in the US and Canada. However, there is no regulatory oversight by either government to ensure proper function of FTS that are being marketed for drug checking. MAIN BODY: Many brands of FTS have rapidly entered the harm reduction market, creating concerns about the reproducibility and accuracy of their performance from brand to brand and lot to lot. Some examples are provided in this Comment. Similar problems with product quality were observed in the mid 2000's when lateral flow immunoassays for malaria were funded in many countries and again in 2020, when COVID-19 tests were in huge demand. The combination of high demand and low levels of regulation and enforcement led some manufacturers to join the goldrush without adequate field testing or quality assurance. We argue that the harm reduction community urgently needs to set a lot checking program in place. A set of simple protocols for conducting the tests and communicating the results have been developed, and are described in the following Perspectives paper in this issue. CONCLUSION: In the absence of governmental regulation and enforcement, the harm reduction community should implement a FTS lot checking program. Based on previous experience with the malaria diagnostic lot checking program, this inexpensive effort could identify products that are not suitable for harm reduction applications and provide valuable feedback to manufacturers. Dissemination of the results will help harm reduction organizations to ensure that FTS they use for drug checking are fit for the purpose.


Subject(s)
Fentanyl , Harm Reduction , Reagent Strips , Humans , Fentanyl/urine , Fentanyl/analysis , Reproducibility of Results , Substance Abuse Detection/methods , Immunoassay/methods , Analgesics, Opioid/urine , Analgesics, Opioid/analysis , COVID-19 , North America
5.
Appl Microbiol Biotechnol ; 108(1): 355, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38822832

ABSTRACT

Getah virus (GETV) is a re-emerging mosquito-borne alphavirus that is highly pathogenic, mainly to pigs and horses. There are no vaccines or treatments available for GETV in swine in China. Therefore, the development of a simple, rapid, specific, and sensitive serological assay for GETV antibodies is essential for the prevention and control of GETV. Current antibody monitoring methods are time-consuming, expensive, and dependent on specialized instrumentation, and these features are not conducive to rapid detection in clinical samples. To address these problem, we developed immunochromatographic test strips (ICTS) using eukaryotically expressed soluble recombinant p62-E1 protein of GETV as a labelled antigen, which has good detection sensitivity and no cross-reactivity with other common porcine virus-positive sera. The ICTS is highly compatible with IFA and ELISA and can be stored for 1 month at 37 °C and for at least 3 months at room temperature. Hence, p62-E1-based ICTS is a rapid, accurate, and convenient method for rapid on-site detection of GETV antibodies. KEY POINTS: • We established a rapid antibody detection method that can monitor GETV infection • We developed colloidal gold test strips with high sensitivity and specificity • The development of colloidal gold test strips will aid in the field serologic detection of GETV.


Subject(s)
Alphavirus , Antibodies, Viral , Gold Colloid , Sensitivity and Specificity , Animals , Gold Colloid/chemistry , Antibodies, Viral/blood , Antibodies, Viral/immunology , Alphavirus/immunology , Swine , Chromatography, Affinity/methods , Alphavirus Infections/diagnosis , Alphavirus Infections/immunology , Swine Diseases/diagnosis , Swine Diseases/virology , Reagent Strips , China , Enzyme-Linked Immunosorbent Assay/methods
6.
J Addict Med ; 18(4): 373-380, 2024.
Article in English | MEDLINE | ID: mdl-38829042

ABSTRACT

BACKGROUND: High potency synthetic opioids like fentanyl have continued to replace or contaminate the supply of illicit drugs in North America, with fentanyl test strips (FTSs) often used as a harm reduction tool for overdose prevention. The available evidence to support FTS for harm reduction has yet to be summarized. METHODS: A search of PubMed, Ovid Embase, and Web of Science was conducted in March 2023. A 2-stage review was conducted to screen by title and abstract and then by full text by 2 reviewers. Data were extracted from each study using a standardized template. RESULTS: A total of 91 articles were included, mostly from North America, predominantly reporting on FTS along with other harm reduction tools, and all conducted after 2016. No randomized controlled trials are reported. Robust evidence exists supporting the sensitivity and specificity of FTS, along with their acceptability and feasibility of use for people who use drugs and as a public health intervention. However, limited research is available on the efficacy of FTS as a harm reduction tool for behavior change, engagement in care, or overdose prevention. CONCLUSIONS: Though FTSs are highly sensitive and specific for point of care testing, further research is needed to assess the association of FTS use with overdose prevention. Differences in FTS efficacy likely exist between people who use opioids and nonopioid drugs, with additional investigation strongly needed. As drug testing with point-of-care immunoassays is embraced for nonfentanyl contaminants such as xylazine and benzodiazepines, increased investment in examining overdose prevention is necessary.


Subject(s)
Drug Overdose , Fentanyl , Harm Reduction , Humans , Fentanyl/urine , Fentanyl/analysis , Drug Overdose/prevention & control , Reagent Strips , Analgesics, Opioid , Opioid-Related Disorders/prevention & control , Illicit Drugs/analysis
7.
Acta Parasitol ; 69(2): 1267-1274, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38753102

ABSTRACT

PURPOSE: This study aimed to develop and evaluate a lateral flow card for the detection of active Schistosoma haematobium infection. METHODS: In order to prepare the immunochromatography lateral flow strip (ICLFS), antibodies purified from schistosomiasis were conjugated passively with gold nanoparticles using a potassium carbonate buffer. RESULTS: The novel ICLFS was able to correctly identify 64 out of 67 samples of schistosomiasis, 6 out of 90 samples of other parasites, and 0 out of 27 control samples. Sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) were 95.5%, 93.3%, 90%, and 91.4% respectively. Comparatively, the sensitivity, specificity, NPV, and PPV of sandwich enzyme-linked immunosorbent assays (ELISA) conjugated with gold nanoparticles (AuNPs) were 91.1%, 88.8%, 85.9%, and 84.4% respectively. The increased sensitivity and specificity of ICLFS produced superior results to those of sandwich ELISA. CONCLUSION: In conclusion, ICLFS is more beneficial and precise than sandwich ELISA for detection of S. haematobium infection at early stage.


Subject(s)
Antigens, Helminth , Chromatography, Affinity , Gold , Metal Nanoparticles , Schistosoma haematobium , Schistosomiasis haematobia , Sensitivity and Specificity , Gold/chemistry , Humans , Schistosoma haematobium/immunology , Metal Nanoparticles/chemistry , Animals , Schistosomiasis haematobia/diagnosis , Chromatography, Affinity/methods , Enzyme-Linked Immunosorbent Assay/methods , Antibodies, Helminth/blood , Reagent Strips
8.
Mikrochim Acta ; 191(6): 325, 2024 05 13.
Article in English | MEDLINE | ID: mdl-38739279

ABSTRACT

Glial fibrillary acidic protein (GFAP) in serum has been shown as a biomarker of traumatic brain injury (TBI) which is a significant global public health concern. Accurate and rapid detection of serum GFAP is critical for TBI diagnosis. In this study, a time-resolved fluorescence immunochromatographic test strip (TRFIS) was proposed for the quantitative detection of serum GFAP. This TRFIS possessed excellent linearity ranging from 0.05 to 2.5 ng/mL for the detection of serum GFAP and displayed good linearity (Y = 598723X + 797198, R2 = 0.99), with the lowest detection limit of 16 pg/mL. This TRFIS allowed for quantitative detection of serum GFAP within 15 min and showed high specificity. The intra-batch coefficient of variation (CV) and the inter-batch CV were both < 4.0%. Additionally, this TRFIS was applied to detect GFAP in the serum samples from healthy donors and patients with cerebral hemorrhage, and the results of TRFIS could efficiently discern the patients with cerebral hemorrhage from the healthy donors. Our developed TRFIS has the characteristics of high sensitivity, high accuracy, and a wide linear range and is suitable for rapid and quantitative determination of serum GFAP on-site.


Subject(s)
Chromatography, Affinity , Glial Fibrillary Acidic Protein , Humans , Biomarkers/blood , Cerebral Hemorrhage/blood , Cerebral Hemorrhage/diagnosis , Chromatography, Affinity/methods , Glial Fibrillary Acidic Protein/blood , Limit of Detection , Reagent Strips
9.
Acta Trop ; 256: 107232, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38729329

ABSTRACT

The World Health Organization calls for schistosomiasis endemic countries to regularly monitor the efficacy of Praziquantel (PZQ) drug, the only antischistosomal drug used for four decades in Tanzania. In response to that call, the current study investigated the efficacy of single dose of PZQ against Schistosoma haematobium during the high transmission season and further assessed, the sensitivity and specificity of urine reagent strips before and after treatment. The study recruited a total of 2,498 -children aged (4 -17 years old) who provided a single urine sample that was visually examined for macro-haematuria, then using urine dipstick and urine filtration technique for microhaematuria and the presence of S. haematobium eggs. The baseline prevalence of S. haematobium eggs positive based on urine filtration test was 29.2 % (95 %CI:27.5-31.0) and that of microhaematuria was 43.1 % (95 %CI:41.1-45.0). Of the infected participants, 40.9 % (95 %CI:37.4-44.6) had a heavy intensity of infection and the geometrical mean intensity (GMI) of infection was 33.7 eggs/10mls of urine. A single dose of PZQ reduced the prevalence of infection to 16.2 %, the GMI of infection to 18.8eggs/10mls of urine and that of microhaematuria to 27.9 %. Cure rate and egg reduction rates (ERR) were 83.8 % and 44.3 % respectively. At baseline, the sensitivity and specificity of the urine reagent strips were 59.7 % and 93.8 %, whereas at post-treatment they were 16.7 % and 93.6 %. When PZQ drug is administered during the high transmission season, its efficacy in term of ERR is poor. The urine reagent strips had low sensitivity but high specificity at pre-and-post PZQ treatment.


Subject(s)
Anthelmintics , Praziquantel , Reagent Strips , Schistosoma haematobium , Schistosomiasis haematobia , Sensitivity and Specificity , Praziquantel/therapeutic use , Praziquantel/administration & dosage , Tanzania/epidemiology , Humans , Schistosomiasis haematobia/drug therapy , Schistosomiasis haematobia/urine , Schistosomiasis haematobia/epidemiology , Child , Animals , Child, Preschool , Female , Male , Anthelmintics/therapeutic use , Anthelmintics/administration & dosage , Schistosoma haematobium/drug effects , Adolescent , Prevalence , Urine/parasitology , Urine/chemistry , Treatment Outcome , Parasite Egg Count
10.
Spectrochim Acta A Mol Biomol Spectrosc ; 317: 124424, 2024 Sep 05.
Article in English | MEDLINE | ID: mdl-38733917

ABSTRACT

A new peptide-based fluorescent probe named DMDH with easy-to-synthesize, excellent stability, good water solubility and large Stokes shift (225 nm) was synthesized for highly selective sequential detections of copper ions (Cu2+) and glyphosate (Glyp). DMDH demonstrated great detection performance towards Cu2+via strong fluorescence quenching, and forming non-fluorescence DMDH-Cu2+ ensemble. As a new promising cascade probe, the fluorescence of DMDH-Cu2+ ensemble was significantly recovered based on displacement approach after glyphosate was added. Interestingly, the limit of detections (LODs) for Cu2+ and glyphosate were 40.6 nM and 10.6 nM, respectively, which were far lower than those recommended by the WHO guidelines for drinking water. More importantly, DMDH was utilized to evaluate Cu2+ and glyphosate content in three real water samples, demonstrating that its effectiveness in water quality monitoring. Additionally, it is worth noting that DMDH was also applied to analyze Cu2+ and glyphosate in living cells in view of significant cells permeability and low cytotoxicity. Moreover, DMDH soaked in filter paper was used to create qualitative test strips and visually identify Cu2+ and glyphosate through significant color changes. Furthermore, smartphone RGB color recognition provided a new method for semi-quantitative testing of Cu2+ and glyphosate in the absence of expensive instruments.


Subject(s)
Copper , Fluorescent Dyes , Glycine , Glyphosate , Peptides , Smartphone , Spectrometry, Fluorescence , Copper/analysis , Copper/chemistry , Glycine/analogs & derivatives , Glycine/analysis , Glycine/chemistry , Fluorescent Dyes/chemistry , Fluorescent Dyes/chemical synthesis , Humans , Spectrometry, Fluorescence/methods , Peptides/chemistry , Limit of Detection , Reagent Strips/analysis , Water Pollutants, Chemical/analysis , HeLa Cells , Drinking Water/analysis
11.
Mikrochim Acta ; 191(6): 311, 2024 05 08.
Article in English | MEDLINE | ID: mdl-38717575

ABSTRACT

Urine retinol-binding protein 4 (RBP4) has recently been reported as a novel earlier biomarker of chronic kidney disease (CKD) which is a global public health problem with high morbidity and mortality. Accurate and rapid detection of urine RBP4 is essential for early monitor of impaired kidney function and prevention of CKD progression. In the present study, we developed a time-resolved fluorescence immunochromatographic test strip (TRFIS) for the quantitative and rapid detection of urine RBP4. This TRFIS possessed excellent linearity ranging from 0.024 to 12.50 ng/mL for the detection of urine RBP4, and displayed a good linearity (Y = 239,581 × X + 617,238, R2 = 0.9902), with the lowest visual detection limit of 0.049 ng/mL. This TRFIS allows for quantitative detection of urine RBP4 within 15 min and shows high specificity. The intra-batch coefficient of variation (CV) and the inter-batch CV were both < 8%, respectively. Additionally, this TRFIS was applied to detect RBP4 in the urine samples from healthy donors and patients with CKD, and the results of TRFIS could efficiently discern the patients with CKD from the healthy donors. The developed TRFIS has the characteristics of high sensitivity, high accuracy, and a wide linear range, and is suitable for rapid and quantitative determination of urine RBP4.


Subject(s)
Chromatography, Affinity , Renal Insufficiency, Chronic , Retinol-Binding Proteins, Plasma , Humans , Retinol-Binding Proteins, Plasma/urine , Chromatography, Affinity/methods , Renal Insufficiency, Chronic/urine , Renal Insufficiency, Chronic/diagnosis , Limit of Detection , Reagent Strips , Biomarkers/urine , Immunoassay/methods
12.
J Immunol Methods ; 530: 113695, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38797275

ABSTRACT

Japanese Encephalitis (JE) is a mosquito borne re-emerging viral zoonotic disease. Sero-conversion in swine occurs 2-3 weeks before human infection, thus swine act as a suitable sentinel for predicting JE outbreaks in humans. The present study was undertaken with the objective of developing immunochromatographic strip (ICS) assay to detect recent infection of Japanese Encephalitis virus (JEV) in swine population. The two formats of ICS assay were standardized. In the first format, gold nanoparticles (GNP) were conjugated with goat anti-pig IgM (50 µg/ml) followed by spotting of recombinant NS1 protein (1 mg/ml) of JEV on NCM as test line and protein G (1 mg/ml) as control line. In the format-II, GNP were conjugated with rNS1 protein (50 µg/ml) followed by spotting of Goat anti-pig IgM (1 mg/ml) as test line and IgG against rNS1 (1 mg/ml) as control line. To decrease the non- specific binding, blocking of serum and nitrocellulose membrane (NCM) was done using 5% SMP in PBS-T and 1% BSA, respectively. Best reaction conditions for the assay were observed when 10 µl of GNP conjugate and 50 µl of 1:10 SMP blocked sera was reacted on BSA blocked NCM followed by reaction time of 15 mins. Samples showing both test and control line were considered positive whereas samples showing only control line were considered negative. A total of 318 field swine sera samples were screened using indirect IgM ELISA and developed ICS assay. Relative diagnostic sensitivity and specificity of format-I was 81.25% and 93.0% whereas of format-II was 87.50% and 62.93%, respectively. Out of 318 samples tested, 32 were positive through IgM ELISA with sero-positivity of 10.06% while sero-positivity with format-I of ICS was 8.1%. Owing to optimal sensitivity and higher specificity of format-I, it was validated in three different labs and the kappa agreement ranged from 0.80 to 1, which signifies excellent repeatability of the developed assay to test field swine sera samples for detecting recent JEV infection.


Subject(s)
Antibodies, Viral , Encephalitis Virus, Japanese , Encephalitis, Japanese , Immunoglobulin M , Metal Nanoparticles , Swine Diseases , Animals , Encephalitis, Japanese/veterinary , Encephalitis, Japanese/diagnosis , Encephalitis, Japanese/immunology , Encephalitis, Japanese/virology , Encephalitis Virus, Japanese/immunology , Swine , Antibodies, Viral/blood , Antibodies, Viral/immunology , Immunoglobulin M/blood , Immunoglobulin M/immunology , Metal Nanoparticles/chemistry , Swine Diseases/diagnosis , Swine Diseases/virology , Swine Diseases/immunology , Swine Diseases/blood , Viral Nonstructural Proteins/immunology , Sensitivity and Specificity , Chromatography, Affinity/methods , Gold/chemistry , Reagent Strips , Reproducibility of Results , Immunoglobulin G/blood , Immunoglobulin G/immunology , Humans
13.
Biosens Bioelectron ; 258: 116357, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38729049

ABSTRACT

The label probe plays a crucial role in enhancing the sensitivity of lateral flow immunoassays. However, conventional fluorescent microspheres (FMs) have limitations due to their short fluorescence lifetime, susceptibility to background fluorescence interference, and inability to facilitate multi-component detection. In this study, carboxylate-modified Eu(III)-chelate-doped polystyrene nanobeads were employed as label probes to construct a multiple time-resolved fluorescent microsphere-based immunochromatographic test strip (TRFM-ICTS). This novel TRFM-ICTS facilitated rapid on-site quantitative detection of three mycotoxins in grains: Aflatoxin B1 (AFB1), Zearalenone (ZEN), and Deoxynivalenol (DON). The limit of detection (LOD) for AFB1, ZEN, and DON were found to be 0.03 ng/g, 0.11 ng/g, and 0.81 ng/g, respectively. Furthermore, the TRFM-ICTS demonstrated a wide detection range for AFB1 (0.05-8.1 ng/g), ZEN (0.125-25 ng/g), and DON (1.0-234 ng/g), while maintaining excellent selectivity. Notably, the test strip exhibited remarkable stability, retaining its detection capability even after storage at 4 °C for over one year. Importantly, the detection of these mycotoxins relied solely on simple manual operations, and with a portable reader, on-site detection could be accomplished within 20 min. This TRFM-ICTS presents a promising solution for sensitive on-site mycotoxin detection, suitable for practical application in various settings due to its sensitivity, accuracy, simplicity, and portability.


Subject(s)
Biosensing Techniques , Edible Grain , Food Contamination , Limit of Detection , Microspheres , Mycotoxins , Zearalenone , Mycotoxins/analysis , Edible Grain/chemistry , Edible Grain/microbiology , Biosensing Techniques/methods , Food Contamination/analysis , Zearalenone/analysis , Chromatography, Affinity/methods , Chromatography, Affinity/instrumentation , Aflatoxin B1/analysis , Aflatoxin B1/isolation & purification , Trichothecenes/analysis , Reagent Strips/analysis , Immunoassay/methods , Immunoassay/instrumentation , Fluorescent Dyes/chemistry
15.
Microb Pathog ; 191: 106669, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38697231

ABSTRACT

African swine fever (ASF) is a lethal disease caused by ASF virus (ASFV), severely impacting the global swine industry. Though nuclear acid-based detection methods are reliable, they are laboratory-dependent. In this study, we developed a device-independent, user friendly and cost-effective quantum dots based immunochromatographic strip (QDs-ICS) with high specificity and sensitivity for the rapid and on-site detection of ASFV antigen. For the preparation of the QDs-ICS, we generated a monoclonal antibody (mAb) mAb-8G8 and polyclonal antibody (pAb) against ASFV-p72 protein. The pAb was labelled with QDs to be used as the detection probe and the mAb-8G8 was coated on the nitrocellulose membrane as the test line. Our results proved that the strip displayed no cross-reactivity with other swine viruses and detection limit of the QDs-ICS was down to 1 ng/mL for the ASFV-p72 protein with great reproducibility. The strip also exhibited high stability with a storage period up to 12 months under room temperature. Twenty blind samples and one hundred clinical samples were examined by the QDs-ICS, conventional PCR and real-time PCR method, respectively. Results showed that the agreement rate between the QDs-ICS and PCR method was 100%, and the agreement rate between the strip and real-time PCR was 94%. The novel QDs-ICS developed here would be an effective tool for on-site detection of ASFV.


Subject(s)
African Swine Fever Virus , African Swine Fever , Antibodies, Monoclonal , Antibodies, Viral , Antigens, Viral , Chromatography, Affinity , Quantum Dots , Sensitivity and Specificity , African Swine Fever Virus/isolation & purification , African Swine Fever Virus/immunology , African Swine Fever Virus/genetics , Animals , African Swine Fever/diagnosis , African Swine Fever/virology , African Swine Fever/immunology , Swine , Antibodies, Monoclonal/immunology , Antibodies, Viral/immunology , Chromatography, Affinity/methods , Antigens, Viral/analysis , Antigens, Viral/immunology , Reproducibility of Results , Reagent Strips
16.
Talanta ; 275: 126181, 2024 Aug 01.
Article in English | MEDLINE | ID: mdl-38692047

ABSTRACT

The detection of biomarkers is of great significance for medical diagnosis, food safety, environmental monitoring, and agriculture. However, bio-detection technology at present often necessitates complex instruments, expensive reagents, specialized expertise, and prolonged procedures, making it challenging to fulfill the demand for rapid, sensitive, user-friendly, and economical testing. In contrast, lateral flow strip (LFS) technology offers simple, fast, and visually accessible detection modality, allowing real-time analysis of clinical specimens, thus finding widespread utility across various domains. Within the realm of LFS, the application of aptamers as molecular recognition probes presents distinct advantages over antibodies, including cost-effectiveness, smaller size, ease of synthesis, and chemical stability. In recent years, aptamer-based LFS has found extensive application in qualitative, semi-quantitative, and quantitative detection across food safety, environmental surveillance, clinical diagnostics, and other domains. This review provided a concise overview of different aptamer screening methodologies, selection strategies, underlying principles, and procedural, elucidating their respective advantages, limitations, and applications. Additionally, we summarized recent strategies and mechanisms for aptamer-based LFS, such as the sandwich and competitive methods. Furthermore, we classified LFSs constructed based on aptamers, considering the rapid advancements in this area, and discussed their applications in biological and chemical detection. Finally, we delved into the current challenges and future directions in the development of aptamer and aptamer-based LFS. Although this review was not thoroughly, it would serve as a valuable reference for understanding the research progress of aptamer-based LFS and aid in the development of new types of aptasensors.


Subject(s)
Aptamers, Nucleotide , Aptamers, Nucleotide/chemistry , Humans , Biosensing Techniques/methods , Reagent Strips/chemistry , SELEX Aptamer Technique/methods , Biomarkers/analysis
17.
Poult Sci ; 103(6): 103648, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38574460

ABSTRACT

Avian infectious bronchitis virus (IBV) still causes serious economic losses in the poultry industry. Currently, there are multiple prevalent genotypes and serotypes of IBVs. It is imperative to develop a new diagnosis method that is fast, sensitive, specific, simple, and broad-spectrum. A monoclonal hybridoma cell, N2D5, against the IBV N protein was obtained after fusion of myeloma SP2/0 cells with spleen cells isolated from the immunized Balb/c mice. The N2D5 monoclonal antibody (mAb) and the previously prepared mouse polyclonal antibody against the IBV N protein were used to target IBV as a colloidal gold-mAb conjugate and a captured antibody, respectively, in order to develop an immunochromatographic strip. The optimal pH and minimum antibody concentration in the reaction system for colloidal gold-mAb N2D5 conjugation were pH 6.5 and 30 µg/mL, respectively. Common avian pathogens were tested to evaluate the specificity of the strip and no cross-reaction was observed. The sensitivity of the strip for detecting IBV was 10-1.4522 EID50/mL. The strip showed a broad-spectrum cross-reactive capacity for detecting IBV antigens, including multiple IBV genotypes in China and all of the seven serotypes of IBV that are currently prevalent in southern China. Additionally, the result can be observed within 2 min without any equipment. The throat and cloacal swab samples of chickens that were artificially infected with three IBV strains were tested using the developed strip and the qPCR method; the strip test demonstrated a high consistency in detecting IBV via qPCR gene detection. In conclusion, the immunochromatographic strip that was established is rapid, sensitive, specific, simple, practical, and broad-spectrum; additionally, it has the potential to serve as an on-site rapid detection method of IBV and can facilitate the surveillance and control of the disease, especially in resource-limited areas.


Subject(s)
Antibodies, Monoclonal , Chickens , Coronavirus Infections , Gold Colloid , Infectious bronchitis virus , Mice, Inbred BALB C , Poultry Diseases , Infectious bronchitis virus/isolation & purification , Infectious bronchitis virus/immunology , Animals , Gold Colloid/chemistry , Poultry Diseases/diagnosis , Poultry Diseases/virology , Coronavirus Infections/veterinary , Coronavirus Infections/diagnosis , Coronavirus Infections/virology , Antibodies, Monoclonal/immunology , Chromatography, Affinity/veterinary , Chromatography, Affinity/methods , Mice , Sensitivity and Specificity , Reagent Strips
19.
Curr Eye Res ; 49(7): 708-716, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38567868

ABSTRACT

PURPOSE: The aim of this article was to investigate whether Schirmer strips gathered during clinical dry eye examinations can be prepared for omics analyses in a standardized way, to adjust for variations in tear volume and enable two separate omics analyses from the same sample. In addition, the intention was to investigate whether fluorescein dye instillation in the eyes gave bias effects on metabolomic analysis. METHODS: Twelve samples from six individuals, with normal or reduced tear production, were collected. Half of the samples were harvested after instillation of fluorescein in the eye. Each strip was divided in half along the length and prepared with a new method for extracting tear content from the Schirmer strip. The new method was established to compensate for different dilutions of metabolites in varying Schirmer strip wetting levels when using identical extraction volume for all samples. Metabolomic data were compared in samples with and without fluorescein dye and Schirmer strips ranging from 1 to 35 mm wetting levels using a global LC-MS method. RESULTS: All samples were successfully analyzed with an average of ∼350 relevant features detected per sample after using both positive and negative electrospray ionization mode, despite low tear volumes in some samples and that only one half of the Schirmer strips were used. Principal component analysis plots and heatmaps revealed no bias effects of fluorescein dye presence or different Schirmer strip values when using the proposed method. CONCLUSION: A high number of relevant metabolomic features can be extracted from longitudinally cut halves of Schirmer strips, which may enable analyses with more than one omics modality from the same sample. With the pre-analytical method described, Schirmer strips can be used for metabolomic analyses even in cases of very low or high tear volume with or without fluorescence.


Subject(s)
Dry Eye Syndromes , Metabolomics , Reagent Strips , Tears , Humans , Tears/chemistry , Tears/metabolism , Metabolomics/methods , Male , Female , Dry Eye Syndromes/metabolism , Dry Eye Syndromes/diagnosis , Adult , Fluorescein/metabolism , Middle Aged , Fluorescent Dyes , Chromatography, Liquid , Spectrometry, Mass, Electrospray Ionization
20.
Molecules ; 29(8)2024 Apr 13.
Article in English | MEDLINE | ID: mdl-38675595

ABSTRACT

The COVID-19 pandemic over recent years has shown a great need for the rapid, low-cost, and on-site detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, an aptamer-based colloidal gold nanoparticle lateral flow test strip was well developed to realize the visual detection of wild-type SARS-CoV-2 spike proteins (SPs) and multiple variants. Under the optimal reaction conditions, a low detection limit of SARS-CoV-2 S proteins of 0.68 nM was acquired, and the actual detection recovery was 83.3% to 108.8% for real-world samples. This suggests a potential tool for the prompt detection of SARS-CoV-2 with good sensitivity and accuracy, and a new method for the development of alternative antibody test strips for the detection of other viral targets.


Subject(s)
Aptamers, Nucleotide , COVID-19 , Gold , Metal Nanoparticles , SARS-CoV-2 , Spike Glycoprotein, Coronavirus , Humans , Aptamers, Nucleotide/chemistry , COVID-19/diagnosis , COVID-19/virology , Gold/chemistry , Limit of Detection , Metal Nanoparticles/chemistry , Reagent Strips , SARS-CoV-2/chemistry , Spike Glycoprotein, Coronavirus/chemistry
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