Colloidal Gold Immunochromatographic Assay Research Articles

Nanobodies-based colloidal gold immunochromatographic assay for specific detection of parathion

Parathion is one of organophosphorus pesticide, which has been prohibited in agricultural products due to its high toxicity to human beings. However, there are still abuse cases for profit in agricultural production. Hence, we established nanobodies-based colloidal gold immunochromatographic assay (GICA) in which nanobodies (Nbs) as an excellent recognition element, greatly improving the stability and sensitivity of ICA. Under the optimal conditions, the developed Nbs-based GICA showed a cut-off value of 50 ng/mL for visual judgment and a half-inhibitory concentration (IC50) of 2.39 ng/mL for quantitative detection. The limit of detection (LOD) was as low as 0.15 ng/mL which was significantly 50-fold higher sensitivity than the commercial mAb-ICA. Additionally, this method exhibited good recoveries for the detection of cabbage, cucumber, and orange samples and excellent correlation with the UPLC-MS/MS method. The results showed that this method developed in this work based on nanobody can be used in practical detection of parathion in foods and nanobody is novel prospective antibody resource for immunoassays of chemical contaminants.

Characterization of viral infections in children with influenza-like-illness during December 2018-January 2019.

Respiratory viral infection (RVI) is of very concern after the outbreak of COVID-19, especially in pediatric departments. Learning pathogen spectrum of RVI in children previous the epidemic of COVID-19 could provide another perspective for understanding RVI under current situation and help to prepare for the post COVID-19 infection control. A nucleic acid sequence-based amplification (NASBA) assay, with 19 pairs of primers targeting various respiratory viruses, was used for multi-pathogen screening of viral infections in children presenting influenza-like illness (ILI) symptoms. Children with ILI at the outpatient department of Beijing Tsinghua Changgung Hospital during the influenza epidemic from 12/2018 to 01/2019 were included. Throat swabs were obtained for both the influenza rapid diagnostic test (IRDT) based on the colloidal gold immunochromatographic assay and the NASBA assay, targeting various respiratory viruses with an integrated chip technology. Of 519 patients, 430 (82.9%) were positive in the NASBA assay. The predominant viral pathogens were influenza A H1N1 pdm1/2009 (pH1N1) (48.4%) and influenza A (H3N2) (18.1%), followed by human metapneumovirus (hMPV) (8.8%) and respiratory syncytial virus (RSV) (6.1%). Of the 320 cases identified with influenza A by NASBA, only 128 (40.0%) were positive in the IRDT. The IRDT missed pH1N1 significantly more frequently than A (H3N2) (P<0.01). Influenza A pH1N1 and A (H3N2) were the major pathogens in <6 years and 6-15 years old individuals respectively (P<0.05). In summary, influenza viruses were the major pathogens in children with ILI during the 2018-2019 winter influenza epidemic, while hMPV and RSV were non-negligible. The coexistence of multiple pathogen leading to respiratory infections is the normalcy in winter ILI cases.

Rapid detection of the freshness of grass carp (Ctenopharygodon idella) fillets by colloidal gold immunochromatography assay

Consumers are greatly concerned about fish freshness. Therefore, this study was aimed to provide a colloidal gold immunochromatography assay (GICA) system for rapid detection of fish freshness. A suitable fish protein was chosen as an antigen for GICA; and the properties and effectiveness of the GICA system were investigated. Results showed that parvalbumin (PV) was suitable as a fish freshness indicator, proven by hierarchical cluster analysis and chosen as a GICA antigen. The GICA system revealed high specificity in the 27–212 μg/L detection range. The correlation coefficient R2 towards enzyme-linked immunosorbent assay reflected high reliability as 0.998. The GICA system was stable within 12 weeks at room temperature. The fitness for indicating freshness by GICA ranged from 87.27% to 111.72%. In short, the GICA system with PV as an antigen could be used as a simple, rapid, and accurate technique for the freshness detection of grass carp fillets.

A rapid colloidal gold immunochromatographic assay based on polyclonal antibodies against HtpsC protein for the detection of Streptococcus suis

An efficient and rapid immunochromatographic assay (ICA) has been engineered for the detection of Streptococcus suis (S. suis). The underpinning principle of this ICA test lies in the use of polyclonal antibodies (pAbs) decorated with colloidal gold, which are specific to S. suis. These pAbs were derived from rabbits immunized with type II histidine triad protein (HtpsC) and HtpsC-N of S. suis. The sensitivity of the ICA was noteworthy, identifying S. suis at bacterial concentrations as diminutive as 1.0 × 103 CFU/mL. Moreover, the assay demonstrated respectable specificity and did not indicate false positives for other bacterial species (Escherichia coli, Salmonella, Staphylococcus aureus, Listeria monocytogenes, Streptococcus pyogenes, Streptococcus lactis, or Enterococcus faecalis). The assay was also capable of detecting multiple S. suis serotypes containing the htpsC gene, including serotypes 1–9, 12, 14, 16 and 23. Nonetheless, the detection of S. suis that lacks the htpsC gene remained beyond the capabilities of this assay. A simultaneous analysis of 16 samples utilizing PCR substantiated the reliability of the ICA test. The assay’s results can be procured within a 15-min window, making it a suitable option for field application. Broadly, this study underscores the potential of the HtpsC protein as a target antigen for the detection of S. suis, and proposes that the HtpsC protein be evaluated further in other detection assays specific for S. suis.

Generating Monoclonal Antibodies against Buprofezin and Developing Immunoassays for Its Residue Detection in Tea Samples.

The synthesis of a hapten and antigen for the preparation of a monoclonal antibody (mAb) for buprofezin is described. The recognition mechanism of hapten and buprofezin by monoclonal antibodies (mAb-19F2) is described. The effectiveness of the mAb-19F2 immunoassay technique was assessed, and the effective detection of buprofezin in tea samples was achieved through the establishment of indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and colloidal gold immunochromatography assay (GICA). The mAb-19F2 subtype was IgG1, with an IC50 of 1.8 ng/mL and a linear range (IC20-IC80) of 0.6-5.4 μg/L, and had a cross-reaction rate of less than 0.18% with 29 other pesticides (neonicotinoids and insect growth regulators). The study identified π-π stacking interactions between hapten and TYR-61 at the mAb-19F2 site and alkyl/phosphate interactions with TRP-105 and ARG-103. The ic-ELISA had an IC50 of 12.9 ng/mL in green tea and 5.65 ng/mL in black tea, with a recovery rate of 92.4%-101.0% and RSD of 2.1%-4.8%. The GICA had a limit of detection (LOD) was 500 ng/mL, with the complete disappearance of the test lines visible to the naked eye. The limit of quantitation (LOQ, IC20) was determined to be 16.8 ng/mL. Additionally, the developed GICA showed no cross-reactivity with neonicotinoid pesticides. The recovery rate of tea spiked recovered samples was 83.6%-92.2%, with an RSD of 5.3%-12.6%, and the results were consistent with the LC/MS method. This study is important for the real-time detection of buprofezin residues to ensure food safety and human health.

Development of multiplex immunochromatographic assay for the simultaneous detection of three polyether ionophore antibiotics in chicken muscle

ABSTRACT A rapid, reliable, sensitive, and multiplex colloidal gold immunochromatographic assay (CGIA) was developed for the simultaneous detection of three polyether ionophore antibiotics (PEs) in chicken muscle. The IC50 values of the optimized CGIA were 1.63, 1.94, and 1.18 ng/mL with a linear range of 0.87–3.04 ng/mL, 0.85–4.41 ng/mL, and 0.57–2.44 ng/mL for maduramicin (MAD), monensin (MON), and salinomycin (SAL), respectively. The cut-off values were 10, 5, and 5 ng/mL, respectively. The whole testing process was completed within 15 min. A parallel analysis in blind chicken muscle samples was conducted by liquid chromatography–tandem mass spectrometry (LC-MS/MS), the developed CGIA did not appear false-positive and false-negative. The results show that the established method could provide a rapid and effective approach for on-site screening and determination of PEs residues in chicken muscle samples.

Effects of rapid drug sensitivity testing for multidrug-resistant bacteria on the prognosis of patients with severe intra-abdominal infection

Objective: To examine the clinical value of rapid detection of drug-resistant bacteria by immunochromatography and the effects of rapid detection on the prognosis of patients with severe intra-abdominal infection complicated by carbapenem-resistant Enterobacteriaceae (CRE) bloodstream infection. Methods: This was a retrospective cohort study. We analyzed clinical data of 73 patients with severe abdominal infections with sepsis or septic shock complicated by CRE bloodstream infection admitted to the general surgery department of Jinling Hospital between February 2022 and February 2023. Patients were divided into a colloidal gold immunochromatographic assay (GICA) group (17 patients) and conventional testing group (56 patients) based on whether a GICA for CRE had been performed on the patients' first blood culture sample during the diagnosis and treatment process. There were no statistically significant differences between the GICA and conventional testing groups in age ([55.9±17.3] vs. [47.6±16.4] years), sex ([16 men vs. one woman ] vs. [41 men vs. 15 women]), median Charlson comorbidity index (3.0[2.0,4.0] vs. 3.0[2.0, 4.8]), septic shock (10 vs. 39), or acute kidney injury (8 vs. 40) (all P>0.05). Both groups routinely underwent traditional bacterial identification and drug susceptibility testing. Additionally, patients in the GICA group were tested directly for positive blood cultures using a GICA carbapenemase test kit. The main outcomes were mortality rates on Days 28 and 90 after the first identification of CRE bloodstream infection in both groups. We also compared the microbial clearance rate, duration of hospitalization and intensive care unit stay, and time from onset of CRE bloodstream infection to initiation of targeted and appropriate antibiotics between the two groups. Results: The rate of microbial clearance of bloodstream infection was significantly greater in the GICA group than in the conventional testing group (15/17 vs. 34/56 [60.7%], χ2=4.476, P=0.034), whereas the 28-day mortality tended to be lower in the GICA than conventional testing group [5/17 vs. 44.6% [25/56], χ2=1.250, P=0.264). The 90-day mortality (8/17 vs. 53.6% [30/56], χ2=0.222, P=0.638), median duration of hospitalization (37.0 [18.0, 46.5] days vs. 45.5 [32.2, 64.8] days, Z=-1.867, P=0.062), and median duration of intensive care unit stay (18.0 [6.5, 35.0] days vs. 32.0 [5.0, 51.8] days, Z=-1.251, P=0.209). The median time between the onset of bloodstream infection and administration of antibiotics was 49.0 (38.0, 69.0) hours in the GICA group, which is significantly shorter than the 163.0 (111.8, 190.0) hours in the conventional testing group (Z=-5.731, P<0.001). The median time between the onset of bloodstream infection and administration of appropriate antibiotics was 40.0 (34.0, 80.0) hours in the GICA group, which is shorter than in the conventional testing group (68.0 [38.2, 118.8]) hours; however, this difference is not statistically significant (Z=-1.686, P=0.093). Conclusions: GICA can provide information on carbapenemase- producing pathogens faster than traditional drug sensitivity testing, enabling early administration of the optimal antibiotics. The strategy of 'carbapenemase detection first' for managing bacterial infection has the potential to improve prognosis of patients and reduce mortality rate.

Molecular epidemiological characterization of hypervirulent carbapenem-resistant Klebsiella pneumoniae in a hospital in Henan Province from 2020 to 2022

Objective: The study investigated the clinical distribution, antimicrobial resistance and epidemiologic characteristics of hypervirulent Carbapenem-resistant Klebsiella pneumoniae (hv-CRKP) in a hospital in Henan Province to provide a scientific basis for antibiotic use and nosocomial infection prevention and control. Methods: A retrospective analysis of the clinical data from the cases was carried out in this study. Clinical data of patients infected with the CRKP strain isolated from the clinical microbiology laboratory of Henan Provincial Hospital of Traditional Chinese Medicine from January 2020 to December 2022 were retrospectively analyzed. A string test, virulence gene screening, serum killing, and a G. mellonella infection model were used to screen hv-CRKP isolates. The clinical characteristics of hv-CRKP and the drug resistance rate of hv-CRKP to twenty-five antibiotics were analyzed using WHONET 5.6. Carbapenemase phenotypic characterization of the hv-CRKP was performed by colloidal gold immunochromatographic assay, and Carbapenemase genotyping, multi-locus sequence typing (MLST) and capsular serotyping of hv-CRKP isolates were performed by PCR and Sanger sequencing. Results: A total of non-duplicate 264 CRKP clinical isolates were detected in the hospital from 2020 to 2022, and 23 hv-CRKP isolates were detected, so the corresponding detection rate of hv-CRKP was 8.71% (23/264). The hv-CRKP isolates in this study were mainly from the intensive care unit (10/23) and neurosurgery department (8/23), and the main sources of hv-CRKP isolates were sputum (10/23) and bronchoalveolar lavage fluid (6/23). The hv-CRKP isolates in this study were highly resistant to β-lactam antibiotics, fluoroquinolones and aminoglycosides, and were only susceptible to colistin, tigecycline and ceftazidime/avibactam. The detection rate of the blaKPC-2 among 23 hv-CRKP isolates was 91.30% (21/23) and none of the class B and class D carbapenemases were detected. Results of MLST and capsular serotypes showed that ST11 type hv-CRKP was the dominant strain in the hospital, accounting for 56.52% (13/23), and K64 (9/13) and KL47 (4/13) were the major capsular serotypes. Conclusion: The hv-CRKP isolates from the hospital are mainly from lower respiratory tract specimens from patients admitted to the intensive care department and the drug resistance is relatively severe. The predominant strains with certain polymorphisms are mainly composed of the KPC-2-producing ST11-K64 and ST11-KL47 hv-CRKP isolates in the hospital.

A colloidal gold test strip based on catalytic hairpin assembly for the clinical detection of influenza a virus nucleic acid

Influenza A epidemics, which occur annually in varying degrees worldwide, is a global challenge to healthcare facilities owing to several limitations of the current detection methods. Therefore, the development of a rapid, convenient, and economical method for the early diagnosis of influenza A will aid clinical treatment and epidemic control. Currently, most of the commonly used clinical rapid tests utilize colloidal gold test strips that detect specific influenza virus antigens but are limited by low sensitivity. Therefore, this study combined catalytic hairpin assembly (CHA) with colloidal gold immunochromatographic assay (GICA) to develop a highly sensitive and visual CHA-GICA test strip. Clinical sample analysis revealed that the sensitivity of the assay was 81.8% and 74% under optimal (35 °C) and room temperature (25 °C) conditions, respectively. In conclusion, this study developed a rapid nucleic acid assay for detecting influenza A virus with high sensitivity and specificity, which can improve the clinical detection of influenza A.

Aflatoxin detection technologies: recent advances and future prospects.

Aflatoxins have posed serious threat to food safety and human health. Therefore, it is important to detect aflatoxins in samples rapidly and accurately. In this review, various technologies to detect aflatoxins in food are discussed, including conventional ones such as thin-layer chromatography (TLC), high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA), colloidal gold immunochromatographic assay (GICA), radioimmunoassay (RIA), fluorescence spectroscopy (FS), as well as emerging ones (e.g., biosensors, molecular imprinting technology, surface plasmon resonance). Critical challenges of these technologies include high cost, complex processing procedures and long processing time, low stability, low repeatability, low accuracy, poor portability, and so on. Critical discussion is provided on the trade-off relationship between detection speed and detection accuracy, as well as the application scenario and sustainability of different technologies. Especially, the prospect of combining different technologies is discussed. Future research is necessary to develop more convenient, more accurate, faster, and cost-effective technologies to detect aflatoxins.

Preparation of a broad-specificity antibody against zearalenone and its primary analogues and development of immunoassay of Coicis Semen and related products.

The purpose of this study was to prepare a highly sensitive and specific zearalenone (ZEN) monoclonal antibody, which was then used to develop an indirect enzyme-linked immunosorbent assay (ic-ELISA) and a colloidal gold immunochromatographic assay (GICA). These techniques were used for the detection of Coicis Semen and related products (Coicis Semen flour, Yimigao, and Yishigao). Immunogens were synthesized by oxime active ester techniques and characterized via ultraviolet spectrophotometry. Immunogens were injected subcutaneously into the abdominal cavities and backs of mice. Using the prepared antibodies, we developed ic-ELISA and GICA rapid detection methods, which were then applied for the rapid detection of ZEN and its analogues from Coicis Semen and related products. For ic-ELISA, the half maximal inhibitory concentration (IC50 ) values for ZEN, α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), zearalanone (ZAN), α-zearalanol (α-ZAL), and β-zearalanol (β-ZAL) were determined to be 1.13, 1.69, 2.06, 0.66, 1.20, and 0.94ng•mL-1 , respectively. For GICA, the cutoff values of ZEN, α-ZEL, β-ZEL, α-ZAL, and β-ZAL on test strips were 0.5ng•mL-1 in phosphate buffer saline (0.01M, pH 7.4), while ZAN was found to be 0.25ng•mL-1 . Furthermore, the cutoff values of test strips were between 10 and 20µg∙kg-1 in Coicis Semen and related products. The results of these two detection methods were in good agreement with results from liquid chromatography-tandem mass spectrometry. This study provides technical support for the preparation of broad-specificity monoclonal antibodies against ZEN and lays the foundation for the simultaneous detection of multiple mycotoxins from food and herbal medicines.

Development of colloidal gold immunochromatography and reverse transcription loop-mediated isothermal amplification assays to detect lychnis mottle virus.

Lychnis mottle virus (LycMoV; genus Unassigned, family Secoviridae) infection of Angelica sinensis produces mottle and mosaic symptoms, damaging the host. Early detection of relevant pathogens is the most critical step in preventing the potential transmission of infectious disease. Polyclonal antibodies (pAbs) with high potency and high specificity were prepared using the recombinant LycMoV capsid protein as an antigen. Here, we developed and optimized a rapid colloidal gold immunochromatography assay (GICA) detection system for LycMoV using this antibody. Under optimum conditions, GICA specifically detected (up to 10,000-fold) positive LycMoV samples. A real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) system was also established by selecting the primers with high sensitivity and specificity to LycMoV. The RT-LAMP detection threshold was 1.42 fg/μl (291 copies/μl). A GICA-RT-LAMP assay system was further established and optimized. The minimum GICA detection line was calculated at 1.52x10-2 ng/μl. Although GICA did not detect positive samples after capturing 2.53x10-3 ng/μl of virus, GICA-LAMP and GICA-RT-PCR did, whose sensitivity was comparatively greater than six-fold. This is the first report showing that GICA-RT-LAMP is a cost effective approach for use in detecting LycMoV without extracting nucleic acids. These sensitive assays will help improve virus disease management in A. sinensis crops.

Rapid detection of apple latent spherical virus infecting Angelica sinensis by gold immunochromatography reverse transcription loop-mediated isothermal amplification assay

Apple latent spherical virus (ALSV; genus: Cheravirus, family: Secoviridae) is capable of infecting Angelica sinensis under natural conditions and causing symptoms of greenish discoloration and mottling, affecting plant growth and causing a reduction in root quality and yield. Virus detection of ALSV is an urgent need for the continued development of angelica industry. Consequently, it is necessary to develop simple, sensitive, and reliable ALSV detection methods. We describe the preparation of polyclonal antibodies with high titer and high specificity using recombinant ALSV coat protein (CP) as the antigen. Using these antibodies, we established and optimized a rapid detection system using colloidal gold immunochromatography assay (GICA) to detect ALSV. A real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) system with a minimum detection line of 2.72 × 103copys/μl was also developed to detect ALSV. This assay was further enhanced using a combination GICA-RT-LAMP assay. This GICA assay successfully detected ALSV in infected plants without cross reactivity recorded from three other plant viruses. The sensitivity of GICA was the same as that of GICA-RT-PCR, whereas the sensitivity of GICA-RT-LAMP was at least 10-fold higher than that of GICA and GICA-RT-PCR. These methods can detect and identify ALSV under field conditions and accurately measure virus concentration in the laboratory.

Sero-prevalence and associated factors of sexually transmitted infections among youth-friendly services Attendees.

Worldwide, more than one million peoples acquire sexually transmitted infections (STIs). The burden of STIs and the youth awareness level on the transmission of STIs is under investigated in Sidama Regional State. To determine the seroprevalence of STIs such as hepatitis B surface antigen (HBsAg), Anti-hepatitis C virus (HCV) antibodies, Human Immunodeficiency virus (HIV) seroprevalence, and syphilis and to determine associated factors among youth-friendly services Attendees at selected health facilities in Hawassa city, Ethiopia. A multicenter cross-sectional study was conducted among 416 randomly selected youth attending youth-friendly services at selected health facilities from May to August 2021. To collect the background characteristics of participants an interviewer-administered questionnaire was used. Blood samples were collected, processed, and tested using Advanced Quality One Step rapid colloidal gold immunochromatographic assay for detection of antibodies for syphilis and hepatitis C virus infection, and hepatitis B virus surface antigen. For the diagnosis of syphilis Rapid Plasma Reagin was also used. HIV1/2 STAT PAK, HIV1/2/O ABON and HIV1/2 SD Bioline were used for testing antibodies for HIV infection. Data entry and analysis were performed using Statistical Package for the Social Sciences (SPSS) version 20.0 software. A crude and adjusted odds ratio with a 95% confidence interval (CI) was computed to identify associated factors. The overall seroprevalence of STIs was 11.5% (48/422), with a 95% CI: (8.7-14.9). Out of the 48 positive results, the proportions of HBsAg, Anti-HCV, HIV, and syphilis were 56.3% (27/48), 27.1% (13/48), 10.4% (5/48), and 6.3% (3/48) respectively. Out of 416 participants, 17.1% responded that it is safe to have sex without using a condom. The Odds of developing STI among female participants, participants who did not identify alcohol intake as a risk factor for STIs, and those who engaged in transactional sex were (AOR = 2.989: 95% CI: 1.27, 7.02), and (AOR = 2.393, 95% CI: 1.18, 4.81) and practice of transactional sex (AOR = 5.527, 95% CI: 1.62, 18.75). STIs are common among youth-friendly services Attendee in Hawassa city The overall STI was significantly associated with sex (females), not able to identify alcohol intake as a risk factor, and practice of transactional sex. High number of (n = 289, 69.5%) participants think that the use of condoms is not effective in preventing STIs and they engage in sexual activity without using condoms.

Development of a colloidal gold immunochromatographic assay strip using monoclonal antibody for rapid detection of porcine deltacoronavirus.

Porcine deltacoronavirus (PDCoV) cause diarrhea and dehydration in newborn piglets and has the potential for cross-species transmission. Rapid and early diagnosis is important for preventing and controlling infectious disease. In this study, two monoclonal antibodies (mAbs) were generated, which could specifically recognize recombinant PDCoV nucleocapsid (rPDCoV-N) protein. A colloidal gold immunochromatographic assay (GICA) strip using these mAbs was developed to detect PDCoV antigens within 15 min. Results showed that the detection limit of the GICA strip developed in this study was 103 TCID50/ml for the suspension of virus-infected cell culture and 0.125 μg/ml for rPDCoV-N protein, respectively. Besides, the GICA strip showed high specificity with no cross-reactivity with other porcine pathogenic viruses. Three hundred and twenty-five fecal samples were detected for PDCoV using the GICA strip and reverse transcription-quantitative real-time PCR (RT-qPCR). The coincidence rate of the GICA strip and RT-qPCR was 96.9%. The GICA strip had a diagnostic sensitivity of 88.9% and diagnostic specificity of 98.5%. The specific and efficient detection by the strip provides a convenient, rapid, easy to use and valuable diagnostic tool for PDCoV under laboratory and field conditions.

Hapten synthesis and a colloidal gold immunochromatographic strip assay to detect nitrofen and bifenox in fruits.

In this study, we synthesized two haptens similar in structure to nitrofen (NIT), and screened out five monoclonal antibodies with the ability to recognize NIT and bifenox (BIF) by competitive ELISA, with the lowest IC50 values of 0.87 ng mL-1 and 0.86 ng mL-1, respectively. The antibody 5G7 was selected to be combined with colloidal gold to establish a lateral flow immunochromatographic assay strip. This method was shown to qualitatively and quantitatively detect the residues of NIT and BIF in fruit samples. The visual limits of detection for qualitative detection were 5 μg kg-1 and 10 μg kg-1 for NIT and BIF, respectively. The calculated limits of detection for quantitative detection were 0.75 μg kg-1, 1.77 μg kg-1 and 2.55 μg kg-1 respectively, for nitrofen in orange, apple and grapes, and 3.54 μg kg-1, 4.96 μg kg-1 and 5.26 μg kg-1, respectively, for bifenox. Thus the strip assay could be used for rapid analysis of fruit samples.

Colloidal Gold Immunochromatography and ELISA Traceability of Tetracycline Residues from Raw Milk to its Dairy Products.

The presence of tetracycline (TC) and its residues in raw milk and milk dairy products poses a threat to human health due to the induction of antibiotic resistance of bacteria that can be transmitted between animals, humans, and the environment. The aim of this study was to investigate the transfer of TC from raw milk to different dairy products: pasteurized milk, boiled milk, sour milk, skimmed milk, and cottage cheese. We analyzed samples of milk from different sources: household farmers, local farms, and milk factories. The analyses of TC in milk and dairy products were performed using colloidal gold immunochromatography assay (GICA) and enzyme-linked immunosorbent assay (ELISA). The highest content of TC was found in the milk purchased from local household farmers; therefore, these samples were chosen for the study of TC transfer to dairy products. TC was also found in sour milk at levels comparable with those obtained in raw milk. The average TC content decreased following heat treatment of the milk, as follows: for pasteurized milk 22.07% and for boiled milk 29.35%. The highest concentrations were determined in cottage cheese in the range 200-620 μg/kg. TC residues are transferred from milk to dairy products in various amounts depending on the preparation conditions, and due to their chemical properties, they accumulate in concentrated derivatives, such as cheese. Therefore, TC can be identified even in cheeses prepared from milk with undetected antibiotic levels.

Monoclonal antibody production and development of immunochromatographic strip assays for screening of the herbicide bispyribac-sodium in rice.

Bispyribac-sodium (BIS) is a new broad-spectrum and efficient herbicide, which is widely used for the control of weeds in rice. To protect the human body from the threat of BIS exposure, it is essential to establish a sensitive and simple detection method. In this work, a high-affinity monoclonal antibody against BIS was produced for the first time, and a colloidal gold immunochromatographic strip assay (ICSA) was developed to screen for BIS in rice samples. The visual limit of detection and the calculated limit of detection of the ICSA were 0.2 μg kg-1 and 0.018 μg kg-1, respectively, which could be accurately obtained within 8 min. The average recoveries of BIS ranged from 90.0% to 109.0% in tests, with CVs ranging from 4.0% to 8.9% for rice samples. Therefore, our ICSA would be a good option for the sensitive and rapid detection of BIS in rice samples.

Diagnostic value of rapid antigen testing for the detection of SARS-CoV-2 infection

Objective: To investigate the diagnostic value of rapid antigen test based on colloidal gold immunochromatographic assay for the detection of SARS-CoV-2 infection in symptomatic patients. Methods: From May 20 to June 5 2022, 76 hospitalized children and their 55 accompanying family members with confirmed SARS-CoV-2 infection in the COVID-19 isolation unit of the Children's Hospital of Fudan University (designated referral hospital for SARS-CoV-2 infection in Shanghai) enrolled. Their nasopharyngeal swab specimens were consecutively collected. The samples were tested for SARS-CoV-2 nucleic acid by real-time quantitative. SARS-CoV-2 antigen was tested by immunochromatography. The correlation between the antigen detection results and the change of the cycle threshold (Ct) values were evaluated, as well as the sensitivity and specificity of SARS-CoV-2 antigen detection at different periods after the onset of the disease. Kappa consistency test was conducted to investigate the consistency between the 2 diagnostic methods. Results: Of the enrolled SARS-CoV-2 symptomatic infections, 76 were children, including 41 males and 35 females, with an age of 5 (2, 9) years; 55 were accompanying families, including 8 males and 47 females, with an age of 38 (32, 41) years. All 478 samples were simultaneously tested for SARS-CoV-2 antigen and nucleic acid. In any period from disease onset to negative conversion of viral nucleic acid, the overall sensitivity of the rapid antigen test was 48.2% (119/247), the specificity was 98.3% (227/231), and antigen test and nucleic acid test showed moderate consistency (κ=0.46, P<0.05). The sensitivity of antigen test was 100% (82/82) when the Ct value was ≤25. And the sensitivity of antigen test was 8/10, 4/15 and 8.3% (3/36) when the Ct value was 26, 30 and 35, respectively. All antigen tests were negative when Ct value was >35. During the period of 1-2 days, 3-5 days, 6-7 days, 8-10 days and >10 days after onset, the sensitivity and specificity of SARS-CoV-2 antigen test were 5/8 and 5/5, 90.2% (37/41) and 5/5, 88.9% (24/27) and 2/5, 45.0% (36/80) and 94.1% (32/34), 18.7% (17/91) and 98.9% (183/185) respectively. The Ct values of nasopharyngeal swabs were<26 during 2 to 7 days after onset, 28.7±5.0 on day 8, 34.5±2.9 on day 13 and > 35 after 14 days, respectively. Conclusion: SARS-CoV-2 antigen test in the patients with SARS-CoV-2 infection shows acceptable sensitivity and specificity within 7 days after onset of disease, and the sensitivity was positively correlated with viral load and negatively correlated with onset time.

Development and Evaluation of a Rapid Neutralizing Antibody Assay for COVID-19 Vaccination.

SARS-CoV-2 neutralizing antibodies have excellent application prospects in the prevention and treatment of COVID-19. This study established a competitive colloidal gold immunochromatography assay (GICA) to detect neutralizing antibodies against the receptor-binding domain (RBD) of SARS-CoV-2 in postvaccination serum. The sensitivity, stability, and specificity of GICA were evaluated using neutralizing antibody solution reference material and positive serum. The consistency and correlation between GICA, pseudovirus neutralization (PN) assay, and ELISA were compared. Consistency analysis of serum neutralizing antibody and specific IgG antibody titers was conducted, and changes in neutralizing antibodies and specific IgG antibodies in serum after inoculation with the homologous booster inactivated vaccine and recombinant vaccine were noted. The sensitivity of the reagent was 20.66 IU/L, and the specificity was 100%. There was a strong consistency and correlation between GICA and PN (κ = 0.886, n = 165; r = 0.918, P < 0.001). The correlation coefficient of serum anti-RBD antibody and specific IgG antibody titers was 0.5253 (P < 0.001). The specific IgG antibody titers in serum after (W4) inoculation with homologous booster inactivated vaccine were 10.80 (S/CO).The anti-RBD antibody titers were 28.33. The anti-RBD omicron variant (B.1.1.529) antibody titers were 11.67. After inoculation with the recombinant vaccine, the specific IgG antibody titers in the serum of W4 were 10.68. The serum anti-RBD antibody titers of W4 were 103.30. The serum anti-RBD omicron variant (B.1.1.529) antibody titers of W4 were 56.67. Therefore, vaccination of the third dose of the homologous booster inactivated vaccine and recombinant vaccine can enhance the level of neutralizing antibodies against the omicron variant (B.1.1.529). This study demonstrates that a GICA kit for neutralizing antibodies against the RBD of SARS-CoV-2 can be used for COVID-19 vaccine evaluation. Changes in titers enable long-term monitoring of a population’s immunity and guide interventions when their immunity declines.