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Dengue fever, caused by the dengue virus and spread mainly by Aedes mosquitoes, is a significant public health concern in tropical regions. Bushehr Province in southern Iran, with its conducive climate for mosquito breeding, shows limited data on dengue incidence, prompting this study. Researchers conducted a study across ten cities in Bushehr, collecting blood samples from 180 participants to detect dengue-specific antibodies through ELISA testing. Results revealed an incidence rate of 2.78%, with positive cases found in Bushehr, Dashtestan, and Tangestan, indicating localized transmission. These findings highlight the risk of dengue outbreaks and emphasize the need for ongoing surveillance, vector control, and public education. Further research is needed to enhance understanding and inform health policies.

Zika virus (ZIKV) is a member of the Flaviviridae family, which includes other clinically notable viruses such as the 4 dengue virus serotypes (DENV-1-4). Distinguishing DENVs from ZIKV using the established serologic assays widely used for monitoring DENV transmission is difficult because of antibody cross-reactivity between these closely related flaviviruses. We describe a modified and improved recombinant envelope domain III-based serologic assay for detecting ZIKV type-specific antibodies in regions with endemic DENV transmission. When the assay was used to measure ZIKV seroprevalence in 2017 among children 9-14 years of age living in a region of the Philippines with endemic DENV transmission, we observed a ZIKV seroprevalence of 18%. Investigators should consider using the ZIKV envelope domain III-based assay, which is simple and readily adaptable for use in standard clinical and public health laboratories, to assess ZIKV seroprevalence in areas with endemic DENV transmission.

The diagnosis of Alzheimer’s disease (AD) is frequently missed or delayed in clinical practice. To remedy this situation, we developed a screening, paper-based (P-ELISA) platform to detect β-amyloid peptide 1–42 (Aβ42) and provide rapid results using a small volume, easily accessible plasma sample instead of cerebrospinal fluid. The protocol outlined herein only requires 3 μL of sample per well and a short operating time (i.e., only 90 min). The detection limit of Aβ42 is 63.04 pg/mL in a buffer system. This P-ELISA-based approach can be used for early, preclinical stage AD screening, including screening for amnestic mild cognitive impairment (MCI) due to AD. It may also be used for treatment and stage monitoring purposes. The implementation of this approach may provide tremendous impact for an afflicted population and may well prompt additional and expanded efforts in both academic and commercial communities.

This chapter contains sections titled: Introduction Use of Immunoassays in Regulatory Systems Implications of Testing for Trade Use in Adhering to Food and Feed Labeling Regulations/Thresholds Global Harmonization Efforts Proficiency Testing International Considerations Certification of Protein‐Based Methods Costs of Testing Conclusions References

This chapter contains sections titled: Introduction to Immunoassay Automation Increasing Throughput by Automation Tissue Collection Sample Processing Sample Transfer Reagent Additions Washing and Reading Steps Analysis Achieving Full Automation by Connecting the Automated Steps Duplexing Future Technologies Summary References

The clinical diagnosis of deep-vein thrombosis (DVT) and pulmonary embolism (PE) is known to be unreli able. Until now, no biological marker has been found to con firm thrombosis, but help can be gained from a biological marker ruling out the diagnosis of DVT or PE, i.e., the sensitive measurement of D-dimer (DD) species. This article summa rizes our experience in introducing a rapid D-dimer test (ELISA VIDAS D-dimères test, bioMérieux) in a collaborative strategy for thrombosis diagnosis during 9 consecutive months involving 1,131 measurements. The efficacy of the DD test was very different according the type of patient, and departments where the DD test provides a real diagnostic benefit were iden tified. High clinical probability for thrombosis was encountered in 32 patients and radiology was carried out, although D-dimer was negative: none of these patients was found to have a throm bosis after radiologic examination. However, extensive prog ress must be made in test prescription to reduce the excessive rate of positive D-dimer tests (78%) and positive measurements that are not followed up by radiology (42%).

We evaluated the diagnostic performance of newly developed microfluidic microplate-based fluorescent ELISA for anti-SARS-CoV-2 antibody detection: the Veri-Q opti COVID-19 IgG and IgM ELISAs (hereafter, “Opti IgG/M”; MiCo BioMed, Gyeonggi-do, Republic of Korea), in comparison with conventional ELISAs. A total of 270 serum samples were analyzed, among which 90 samples were serially obtained from 25 COVID-19 patients. Another 180 samples were collected from 180 SARS-CoV-2–negative individuals. As comparative assays, we used SCoV-2 Detect IgG/M ELISA (hereafter, “InBios IgG/M”; InBios, Seattle, WA, USA) and Veri-Q COVID-19 IgG/IgM ELISA (hereafter, “Veri-Q IgG/M”; MiCo BioMed). Compared with conventional ELISAs, the Opti IgG yielded 97.1–100.0% positive percent agreement, 95.2–98.0% negative percent agreement, 96.3–97.8% total percent agreement, and kappa values of 0.90–0.94. Between the Opti IgM and the InBios IgM, the values were 93.7%, 96.6%, 95.9%, and 0.89, respectively. For the Opti IgG, sensitivities for the samples collected from 0–7, 8–14, 15–21, and ≥ 22 days after symptom onset were 40.0, 58.3, 94.1, and 100.0%, respectively. The values for the Opti IgM were 30.0, 54.2, 88.2, and 80%, respectively. The diagnostic specificities of the Opti IgG and IgM were 99.4 and 97.2%, respectively. The microfluidic microplate-based fluorescent ELISAs showed comparable diagnostic performance to conventional ELISAs for detecting anti-SARS-CoV-2 antibodies. With the combination of high throughput, a simplified workflow, and the ability to analyze reduced volumes, this new technology has great potential for improving SARS-CoV-2 serologic testing.

The accurate and rapid detection of H1N1 viral specific protein is crucial for effective disease control and management. Traditional diagnostic methods based on ELISA immunoassay often require complex manual operations, long incubation times, and lack the integration needed for automated, high-throughput analysis. There is an urgent demand for a portable, sensitive, integrated, and fully automated diagnostic platform to address these challenges in medical diagnostics. We developed an automated diagnostic system that integrates nanozyme-based detection within a centrifugal microfluidic device for rapid H1N1 viral specific protein detection. The system utilizes nanozymes for signal amplification, leveraging nanozymes to reduce assay times. A high-resolution image sensor and image processing algorithm enable precise interrogation of test results, demonstrating significant improvements in sensitivity. The compact centrifugal microfluidic device automates the sequential delivery of samples, reagents, and nanozyme probes, seamlessly implements mixing, incubation, and liquid transfer. The system achieves a 50-fold sensitivity improvement over the conventional method based on colloidal gold lateral flow strips, with a test time of only 15 min. This work develops a fully automated, integrated diagnostic platform by combining nanozyme-enhanced immunoassay with automatic centrifugal microfluidics. The platform eliminates manual intervention, achieving high sensitivity and rapid detection of infectious disease viral specific protein. Its compact and integrated design makes it a versatile tool for point-of-care diagnostics.

The enzyme-linked Immunosorbent Assay (ELISA) is the gold standard clinical diagnostic tool for the detection and quantification of protein biomarkers. However, conventional ELISA tests have drawbacks in their requirement of time, expensive equipment and expertise for operation. Hence, for the purpose of rapid, high throughput screening and point-of-care diagnosis, researchers are miniaturizing sandwich ELISA procedures on Lab-on-a-Chip and Lab-on-Compact Disc (LOCD) platforms. This paper presents a novel integrated device to detect and interpret the ELISA test results on a LOCD platform. The system applies absorption spectrophotometry to measure the absorbance (optical density) of the sample using a monochromatic light source and optical sensor. The device performs automated analysis of the results and presents absorbance values and diagnostic test results via a graphical display or via Bluetooth to a smartphone platform which also acts as controller of the device. The efficacy of the device was evaluated by performing dengue antibody IgG ELISA on 64 hospitalized patients suspected of dengue. The results demonstrate high accuracy of the device, with 95% sensitivity and 100% specificity in detection when compared with gold standard commercial ELISA microplate readers. This sensor platform represents a significant step towards establishing ELISA as a rapid, inexpensive and automatic testing method for the purpose of point-of-care-testing (POCT) in resource-limited settings.

Foodborne pathogens have been a cause of a large number of diseases worldwide and more so in developing countries. This has a major economic impact. It is important to contain them, and to do so, early detection is very crucial. Detection and diagnostics relied on culture-based methods to begin with and have developed in the recent past parallel to the developments towards immunological methods such as enzyme-linked immunosorbent assays (ELISA) and molecular biology-based methods such as polymerase chain reaction (PCR). The aim has always been to find a rapid, sensitive, specific and cost-effective method. Ranging from culturing of microbes to the futuristic biosensor technology, the methods have had this common goal. This review summarizes the recent trends and brings together methods that have been developed over the years.