Saliva Antigens Research Articles

Mysterious Allergy Caused by Tick Bite: Alpha-Gal Syndrome.

Alpha-Gal syndrome (AGS) manifests as an intricate allergic response characterised by the formation of specific immunoglobulin E (IgE) antibodies targeting a carbohydrate termed galactose-a-1.3-galactose (a-Gal). Alpha-Gal antigens, which play a role in AGS, have been detected in the salivary glands and saliva of various tick species, especially Amblyomma americanum. Identifying these antigens in tick saliva underlines the potential role of tick bites in sensitising individuals to a-Gal and contributes to the complex immunological processes associated with AGS. When people with a-Gal allergy eat beef, pork, lamb, or the flesh of other mammals, they experience an allergic reaction that causes various symptoms, including rash, nausea, vomiting, and diarrhoea. In some cases, AGS can be life-threatening requiring emergency medical attention. Moreover, these reactions do not occur only due to red meat; intake of medical drugs, vaccines, and antidotes containing a-Gal epitopes can also trigger allergies. The fact that the symptoms causing IgE antibodies are directed against a carbohydrate moiety the unusual delay between food consumption and the onset of symptoms, and the differences in the reactions shown by a-Gal allergy make a-Gal syndrome an unprecedented allergic disease and distinguish it from other food allergies. Interestingly, a-Gal antigens involved in the development of AGS have been discovered in salivary secretions of different tick species in several continents. However, the underlying causes of a-Gal-specific IgE production and immune responses to tick bites are not fully understood. This complex system is crucial for identifying and developing new therapies for the disease. This article reviews the evolution of a-Gal, the current understanding of AGS and its relationship to tick species.

Genetic and mechanistic evaluation of an individual with para-Bombay phenotype associated with a compound heterozygote comprising two novel FUT1 variants.

As is well documented, the para-Bombay phenotype is typically characterized by the reduction or absence of ABH antigens on red blood cells but the presence of corresponding antigens in saliva. Herein, the underlying molecular mechanism of an individual with para-Bombay AB phenotype combined with two novel variants of the FUT1 gene was investigated. ABH antigens and antibodies were detected in the serum of the proband using conventional serological methods. The coding region nucleotides of the ABO, FUT1, and FUT2 genes were directly sequenced by polymerase chain reaction. Moreover, the FUT1 haploid type in the proband was analyzed by TA clone sequencing. The 3D structure of wild-type and mutant fucosyltransferases were simulated and analyzed using Phyre2 and Pymol software. Lastly, the effect of missense substitution on the function of fucosyltransferase was predicted by the Polymorphism Phenotyping algorithm (PolyPhen-2) and MutationTaster. ABH antigens were noted to be absent on the surface of red blood cells of the proband. The ABO genotype was ABO*A1.02/ABO*B.01, while the FUT2 genotype was FUT2*01/FUT2*c.357T. Interestingly, two novel missense variants (c.289G>A, p.Ala97Thr and c.575G>C, p.Arg192Pro) and one synonymous SNP (c.840G>A) were identified in the FUT1 gene. Furthermore, c.289G>A was detected in one haploid type, whereas c.575G>C and c.840G>A were discovered in another haploid type. Meanwhile, in silico analysis revealed that amino acid substitution caused by missense variants altered the partial spatial structure of the α-helices where residues 97 and 298 were located using 3D homology modeling software. Finally, both missense variants were defined as probably damaging based on PolyPhen-2 prediction. Two novel FUT1 variants were identified in a Chinese individual with para-Bombay AB phenotype, which can expand our understanding of the molecular mechanism underlying the para-Bombay phenotype and contribute to improving the safety of blood transfusion.

A modified gate oxide tunnel Field-Effect transistor (TFET) biosensor to identify receptor Tyrosine-Protein kinase 2 (C-erbB-2) in Serum/Saliva

This research demonstrates an analysis of a modified gate oxide tunnel field-effect transistor (TFET) with pocket (Poc-MGOTFET) biosensor for the incredibly sensitive and real-time identification of the breast cancer (BC) biomarker receptor tyrosine-protein kinase (C-erbB-2) in serum and saliva. This research investigates the impact of interface charge modulation in Poc-MGOTFET biosensor with embedded cavities for rapid, accurate, and reliable identification of antigens in bodily fluids, like serum and saliva. The TCAD Sentaurus is used to numerically simulate the proposed biosensor in 2D. The dual cavity etched underneath the dual gate electrode of the proposed biosensor facilitates biomolecule immobilization. This enhances the ability of biomolecules to regulate the source/channel tunneling rate and the proposed biosensor’s electrical performance metrics. A numerical model is developed for the interface charge equivalent of the C-erbB-2. The sensitivity of the device to different C-erbB-2 concentrations in serum/saliva has been studied. The results of our research demonstrate that Poc-MGOTFET, featuring an extended gate structure, modified gate oxide, and pocket along the source side, reduces the leakage current (ILeakage) and OFF current (IOFF), enhances the probability of tunneling, boosts gate control for a greater ON current (ION) and ION/IOFF ratio, and increases sensitivity. Moreover, increased sensitivity by an order of magnitude 106 results from the effect of interface charges corresponding to varying concentrations of C-erbB-2 biomarkers on the sensitivity of the biosensor (as measured by the ION/IOFF ratio).

A Multiplex "Disposable Photonics" Biosensor Platform and Its Application to Antibody Profiling in Upper Respiratory Disease.

Photonic technologies promise to deliver quantitative, multiplex, and inexpensive medical diagnostic platforms by leveraging the highly scalable processes developed for the fabrication of semiconductor microchips. However, in practice, the affordability of these platforms is limited by complex and expensive sample handling and optical alignment. We previously reported the development of a disposable photonic assay that incorporates inexpensive plastic micropillar microfluidic cards for sample delivery. That system as developed was limited to singleplex assays due to its optical configuration. To enable multiplexing, we report a new approach addressing multiplex light I/O, in which the outputs of individual grating couplers on a photonic chip are mapped to fibers in a fiber bundle. As demonstrated in the context of detecting antibody responses to influenza and SARS-CoV-2 antigens in human serum and saliva, this enables multiplexing in an inexpensive, disposable, and compact format.

Modified streptavidin–biotin based lateral flow test strip for rapid detection of SARS-CoV-2 S1 antigen in saliva samples

Compared to other infectious diseases, for which LFT development can take years, SARS-CoV-2 antigen LFTS were developed and deployed within months. LFTS for antigen detection were adopted on an unprecedented scale during the COVID-19 pandemic, but many of them lack the sensitivity especially for samples with low viral load. In our previous work, we developed an enhanced signal strip for detection of SARS CoV-2 SI antigens in saliva. Here we introduce some modification to improve the sensitivity, and specificity, and to lower the cost of the strip, by using biotin streptavidin (BS) system. In the modified BS strip, gold-streptavidin and biotinylated Nanobodies (Nbs) against S1 antigen were externally mixed with the tested samples (saliva or nasopharyngeal swab) before their application on the sample pad of the test strip containing angiotensin converting enzyme (ACE-2), as the capturing probe. The study included 320 individuals, with 180 being positively confirmed by RT-PCR and 140 confirmed negative, as well as, 45 health care workers, who were responsible for screening and handling of surgical cases in General Surgery Department and COVID clinic of TBRI. Our results proved that modified BS strip improved the overall sensitivity and specificity of S1antigen detection in saliva samples (95.21% and 99.29% respectively) compared to our previously developed enhanced LFTS (91.66% and 98.57% respectively). Also, the sensitivity of cases with Ct ≤ 30, Ct ≤\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\le$$\\end{document} 35, and Ct ≤\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\le$$\\end{document} 40 using the modified BS strip showed higher values (98.54%, 95.38%, and 88.89% respectively), compared to the corresponding results of our previously developed enhanced LFTS (95.86%, 92.31%, and 82.22% respectively). There were no cross-reactions with either Middle East respiratory syndrome corona virus MERS-CoV or SARS-CoV antigens. Furthermore, we found that the lower viral detection limit (LVD) of BS strip was obviously lower than our previous LVD limit of the enhanced LFTS (0.2 × 104 copies/ml vs. 0.4 × 104 copies/ml, respectively). Our developed BS strip showed that saliva samples gave better results than nasopharyngeal swabs of the same patients. The fact of using smaller amounts of Nbs, and ACE2, as well as the dispensing off of conjugate pad when applying BS strip modifications, justified the expected reduction in the costs of the strip. The implementation of BS strips on saliva samples of 45 health co-workers, who were tested 4 and 6 days after exposure to infection, showed an increase in the sensitivity, starting from the 4th day and reaching its highest level on the 6th day in both high risk and paramedic groups (90.9%, and 80.0%, respectively). This study provides evidence that employment of the modified BS system could increase the sensitivity of the strips, lower their cost, and render them an effective screening tool for early detection of the virus in saliva of suspected Covid-19 patients.

An Analytical Protocol for Detecting Antibody Titer Levels in Serum/Saliva by Indirect Enzyme-Linked Immunosorbent Assay (ELISA).

Enzyme-linked immunosorbent assay (ELISA) detects qualitatively and quantitatively the presence of antibodies or antigens in a sample. Due to its simplicity, high sensitivity, and user-friendliness, the test is widely used in laboratory research, clinical diagnoses, and food testing. This chapter describes the indirect semiquantitative ELISA protocol used to monitor antibody levels in animals and analyze the titer levels of specific antibodies against a target antigen in serum and saliva.

Prostate-specific antigen: An unfamiliar protein in the human salivary glands.

The presence of prostate-specific antigen (PSA) in saliva and salivary glands has been reported. Nevertheless, its release pathway in these glands remains to be elucidated. Here, we showed PSA subcellular distribution focusing on its plausible route in human salivary parenchyma. Sections of parotid and submandibular glands were subjected to the immunohistochemical demonstration of PSA by the streptavidin-biotin method revealed by alkaline phosphatase. Moreover, ultrathin sections were collected on nickel grids and processed for immunocytochemical analysis, to visualize the intracellular distribution pattern of PSA through the observation by transmission electron microscopy. By immunohistochemistry, in both parotid and submandibular glands PSA expression was detected in serous secretory acini and striated ducts. By immunocytochemistry, immunoreactivity was retrieved in the cytoplasmic compartment of acinar and ductal cells, often associated with small cytoplasmic vesicles. PSA labeling appeared also on rough endoplasmic reticulum and in the acini's lumen. A negligible PSA labeling appeared in most of the secretory granules of both glands. Our findings clearly support that human parotid and submandibular glands are involved in PSA secretion. Moreover, based on the immunoreactivity pattern, its release in oral cavity would probably occur by minor regulated secretory or constitutive-like secretory pathways.

Electrochemical Paper-Based Nanobiosensor for Rapid and Sensitive Detection of Monkeypox Virus.

Monkeypox virus (MPXV) infection was classified as a public health emergency of international concern by the World Health Organization (WHO) in 2022, being transmitted between humans by large respiratory droplets, in contact with skin lesions, fomites, and sexually. Currently, there are no available accessible and simple-to-use diagnostic tests that accurately detect MPXV antigens for decentralized and frequent testing. Here, we report an electrochemical biosensor to detect MPXV antigens in saliva and plasma samples within 15 min using accessible materials. The electrochemical system was manufactured onto a paper substrate engraved by a CO2 laser machine, modified with gold nanostructures (AuNS) and a monoclonal antibody, enabling sensitive detection of A29 viral protein. The diagnostic test is based on the use of electrochemical impedance spectroscopy (EIS) and can be run by a miniaturized potentiostat connected to a smartphone. The impedimetric biosensing method presented excellent analytical parameters, enabling the detection of A29 glycoprotein in the concentration ranging from 1 × 10-14 to 1 × 10-7 g mL-1, with a limit of detection (LOD) of 3.0 × 10-16 g mL-1. Furthermore, it enabled the detection of MPXV antigens in the concentration ranging from 1 × 10-1 to 1 × 104 PFU mL-1, with an LOD of 7.8 × 10-3 PFU mL-1. Importantly, no cross-reactivity was observed when our device was tested in the presence of other poxvirus and nonpoxvirus strains, indicating the adequate selectivity of our nanobiosensor for MPXV detection. Collectively, the nanobiosensor presents high greenness metrics associated with the use of a reproducible and large-scale fabrication method, an accessible and sustainable paper substrate, and a low volume of sample (2.5 μL), which could facilitate frequent testing of MPXV at point-of-care (POC).

Hydrogel‐Gated Silicon Nanotransistors for SARS‐CoV‐2 Antigen Detection in Physiological Ionic Strength

AbstractThe recent Coronavirus Disease 2019 (COVID‐19) outbreak strongly propels advancements in biosensor technology, leading to the emergence of novel methods for virus detection. Among them, those using nanostructured field‐effect transistors (FETs) provide an ultrasensitive approach toward point‐of‐care diagnostics. However, the application of these biosensors in analyzing biofluids has been limited by their reduced screening length in high ionic strength liquids. To address this challenge, a solution is presented involving the surface modification of FETs with a hydrogel based on star‐shaped polyethylene glycol. This hydrogel is loaded with specific antibodies against the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) spike protein. By incorporating the hydrogel, the effective Debye length is effectively increased, thereby preserving the sensitivity in biofluids. The efficacy of this approach is demonstrated by employing silicon nanonet‐based FETs for the detection of viral antigens in both buffer and saliva, as well as cultured viral particle dispersions. Moreover, positive and negative patient samples are successfully differentiated, showcasing the practical application of this method. Finally, a theoretical frame is proposed to elucidate the underlying mechanism behind the preservation of sensitivity.

Interfacial Polymer Engineered Field Effect Transistor Biosensors for Rapid and Efficient Identification of SARS‐CoV‐2 N Antigen

Comprehensive SummaryWith the advantages of high sensitivity, rapid response, label‐free, and simple operation, field effect transistor biosensors have shown promising application prospects in large‐scale pathogen screening. However, in practical biological fluids with relatively high ionic strength, such as saliva and serum, the Debye screening effect will weaken the interaction between FET biosensors and target bio‐molecules, thereby affecting the sensing sensitivity and accuracy. Herein, an interfacial polymer‐engineered field effect transistor (IPE FET) biosensor was developed for the efficient identification of SARS‐CoV‐2 N antigens in saliva samples. The inclusion of a polymer layer shortens the distance between target molecules and the electrode interface, which effectively overcomes the limitation of Debye screening. The constructed IPE FET biosensors exhibit high sensitivity and anti‐fouling capability, achieving efficient detection of SARS‐CoV‐2 N antigen in saliva within 5 min with a detection limit of 4.6 fg·mL−1. In a cohort of 35 simulated throat swab samples of SARS‐CoV‐2 N antigen, IPE FET exhibits an identification accuracy of up to 97.1%, with predictive sensitivity and specificity of 96.0% and 100.0%, respectively. The excellent performance of IPE FET not only provides a strategy to design efficient detection platforms but also suggests a pathway to realize rapid and scalable epidemic screening.

Opal photonic crystal-enhanced upconversion turn-off fluorescent immunoassay for salivary CEA with oral cancer.

The point-of-care test of tumor markers in saliva with high specificity and sensitivity for early diagnosis of oral cancer is of great interest and significance, but remaining a daunting challenge due to the low concentration of such biomarkers in oral fluid. Herein, a turn-off biosensor based on opal photonic crystal (OPC) enhanced upconversion fluorescence is proposed to detect the carcinoembryonic antigen (CEA) in saliva by applying fluorescence resonance energy transfer sensing strategy. Hydrophilic PEI ligands are modified on upconversion nanoparticles to enhance the sensitivity of biosensor by promoting sufficient contact between saliva and detection region. As a substrate for the biosensor, OPC can also provide a local-field effect for greatly enhanced upconversion fluorescence by coupling the stop band and excitation light, and a 66-fold amplification of the upconversion fluorescence signal was obtained. For the CEA detection in spiked saliva, such sensors showed a favorable linear relationship at 0.1-2.5ngmL-1 and more than 2.5ngmL-1, respectively. The limit of detection was down to 0.1ngmL-1. Moreover, by monitoring real saliva, the effective discrepancy between patients and healthy people was confirmed, indicating remarkable practical application value in clinical early diagnosis and home-based self-monitoring of tumors.

Self-enhancement lateral flow immunoassay for COVID-19 diagnosis

Equipment-free colorimetric-based lateral flow immunoassay (LFIA) is the most convenient and popular tool for various applications, including diagnostic tools requiring high sensitivity for the detection of pathogens. Thus, improvements and developments of LFIA are constantly being reported. Herein, we enriched the sensitivity of LFIA using the gold enhancement principle, emphasizing needlessly complicated apparatus, only one step for the strip test operation, and typical time incubation (15 min) process. Self-enhanced LFIA was then executed for subsequent flows by overlapping the additionally enhanced pad composed of gold ions and reducing agent on the conjugate pad and the sample pad. Self-enhanced LFIA was performed to detect SARS-CoV-2 antigens in saliva. The obtained result depicted that the achieved sensitivity was up to tenfold compared with that of conventional LFIA by visual measurements. The detection limits of self-enhanced LFIA detecting nucleocapsid protein antigens in the saliva sample was 0.50 and 0.10 ng/mL employed by naked eye detection and calibration curve-based calculation, respectively. When the proposed device was applied to 207 human saliva samples, the diagnostic performance presented a 96.10 % sensitivity and 99.23 % specificity. This self-enhanced LFIA could be implemented in large-scale production and demonstrates higher sensitivity with effortless use, which meets the requirements for point-of-care testing and on-field mass screening.

Assessment of Salivary ABO Blood Group Antigens and Secretor Status in Sriganganagar, Rajasthan: A Correlational Analysis of 300 Samples.

Aim To estimate the ABO blood groups from saliva samples and to correlate with the secretor status. Materials and methods A sample size of 300 individuals was selected from the outpatient department of Surendera Dental College & Research Institute, Sriganganagar, India, and from dental camps organized by the college in the near vicinity. Informed consent was obtained from selected individuals to collect their blood and saliva samples. Salivary samples were evaluated for ABO blood groups by the absorption-inhibition method. The indicator erythrocytes were prepared after blood group confirmation from serum. It was used to identify the blood group antigens in saliva to confirm the secretor status. The results were tabulated and the Pearson's chi-squared test was performed for statistical analysis using SPSS 15.0 (SPSS Inc., Chicago, IL). Results The present study showed that 282 subjects (94%) were Rhesus positive and 18 subjects (6%) were Rhesus negative. Two-hundred-and-fifty subjects (83.3%) were secretors of antigens in saliva. Non-secretors were 50 subjects (16.7%). We identified that 250/300 were secretors and the majority were in AB & A group. Conclusion Blood groups could not be detected from the saliva of subjects who were non-secretors. In contrast, blood types could be accurately identified from the saliva of those subjects who were secretors of antigen.

Use of Zebrafish Embryos To Reproduce Human Norovirus and To Evaluate Human Norovirus Infectivity Decay after UV Treatment.

This study reports an essential improvement of the method for replication of human norovirus (HNoV) with the use of zebrafish (Danio rerio) embryos. With three HNoV genotypes and P-types GII.2[P16], GII.4[P16], and GII.17[P31], we demonstrated that this tool had higher efficiency and robustness than the zebrafish larvae as reported previously. When zebrafish larvae were injected with virus (1.6 ± 0.3 log genome copies/10 larvae), a significant increase of virus genome copies was detected at 2 days postinfection (dpi; 4.4 ± 0.8 log genome copies/10 larvae, P < 0.05) and the viral loads started to decrease gradually from 3 dpi. In comparison, when the viruses were injected into the zebrafish embryos, significant virus replication was noticed from 1 dpi and lasted to 6 dpi (P < 0.05). The virus levels detected at 3 dpi had the highest mean value and the smallest variation (7.7 ± 0.2 log genome copies/10 larvae). The high levels of virus replication enabled continuous passaging for all three strains up to four passages. The zebrafish embryo-generated HNoVs showed clear patterns of binding to human histo-blood group antigens (HBGAs) in human saliva by a simple saliva-binding reverse transcription-quantitative PCR (RT-qPCR). Last, in a disinfection study, it was shown that a dose of 6 mJ/cm2 UV254 was able induce a >2-log reduction in HNoV infectivity for all three HNoV strains tested, suggesting that HNoVs were more UV susceptible than multiple enteric viruses and commonly used HNoV surrogates as tested before. IMPORTANCE HNoVs are a leading cause of gastroenteritis outbreaks worldwide. The zebrafish embryo tool as developed in this study serves as an efficient way to generate viruses with high titers and clean background and a straightforward platform to evaluate HNoV inactivation efficacies. It is expected that this tool will not only benefit epidemiological research on HNoV but also be used to generate HNoV inactivation parameters which are highly needed by the water treatment and food industries.

Saliva and wastewater surveillance for SARS-CoV-2 during school reopening amid COVID-19 pandemic in Thailand

ObjectivesSchool closure during the coronavirus disease 2019 (COVID-19) pandemic resulted in a negative impact on children. Serial testing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been proposed as a measure for safety school reopening. We aimed to study the usefulness of SARS-CoV-2 surveillance by saliva testing and performing wastewater surveillance for SARS-CoV-2 in a day school in a resource-limited setting. MethodsWe conducted a cluster randomized study to investigate the potential use of saliva antigen testing compared to saliva pooling for nucleic acid detection in a primary school in Thailand from December 2021 to March 2022. Wastewater surveillance in the school was also performed. ResultsA total of 484 participants attended the study. SARS-CoV-2 was detected in two participants from the tests provided by the study (one in the pool nucleic acid test arm, and another in the quantitative antigen test arm). Additional ten participants reported positive results on an additional rapid antigen test (RAT) performed by nasal swab when they had symptoms or household contact. There was no difference among arms in viral detection by intention-to-treat and per protocol analysis (p = 0.304 and 0.894, respectively). We also investigated the feasibility of wastewater surveillance to detect the virus in this setting. However, wastewater surveillance could not detect the virus. ConclusionsIn a low COVID-19 prevalence, serial saliva testing and wastewater surveillance for SARS-CoV-2 rarely detected the virus in a day school setting. Performing RAT on nasal swabs when students, teachers or staff have symptoms or household contact might be more reasonable.

Carbohydrate larval antigen (CarLA IgA) responses to mixed species nematode infection in pasture grazed Angora goats

The genetics of indicator traits for resistance of Angora goats to gastrointestinal nematode parasite infections, and their relationships with productivity traits, were investigated on a commercial mixed-enterprise farm in the eastern North Island of New Zealand. Faecal egg counts (FEC), specific Immunoglobulin A (IgA) and Immunoglobulin G (IgG) antibody titres against carbohydrate larval antigen (CarLA) in saliva, live weight and fleece weights were recorded from 278 goats of 19–20 months of age, run as four separate mobs (breeding bucks, castrated males (wethers), or 2 groups of breeding does). Summary statistics showed the mobs differed significantly in liveweight, loge (FEC+50), loge (IgA) and loge (IgG). Genetic parameters were estimated using an animal model with repeated records where appropriate, after adjusting for the different contemporary animal groups, using the restricted maximum likelihood (REML) package. Heritability estimates from repeated measures were 0.19 ± 0.16 for FEC, 0.28 ± 0.16 for CarLA specific IgA and 0.23 ± 0.15 for CarLA specific IgG. The CarLA specific IgA response was negatively genetically correlated with FEC (−0.99 ± 0.31) suggesting that it could be used as a selection tool for breeding resistant animals. Although the genetic and phenotypic correlations between CarLA IgA and IgG were high and significant, the analysis between loge (FEC+50) and loge CarLA IgG did not converge. Further, both FEC and CarLA IgA showed significant and favourable genetic correlations with live weight. In contrast, CarLA IgG showed an unfavourable phenotypic correlation with liveweight. While this is only a preliminary study, the results do suggest that the immunoassay measuring salivary CarLA IgA response may have utility as a selection tool for parasite resistance in some breeds of goats.

A Comparison of Absorption Inhibition and Absorption Elution Techniques for Detection of ABO Blood Groups in Saliva in Meitei Population: A Cross Sectional Study

The ABO blood group system is the significant element for forensic serological examination of blood and body fluids. ABO blood groups are the primary, most common, conspicuous, and easily detectable groups. These blood group specific antigens are abundantly present in many other bodily secretions such as sweat, semen and even saliva. Many studies have detected the presence of ABO blood groups in saliva by using both the absorption inhibition and absorption elution methods. This study of blood group ABO antigens in saliva was carried out using both the techniques in a tertiary care centre in Imphal, Manipur. Using absorption elution method, out of the 90 secretors subjects, positive results were obtained in 82 (91.1%) subjects and using absorption inhibition method, positive results were detected in 71 (78.9%) subjects. On statistical analysis, the mean age in completed years of the participants is calculated as 22.12 years with a standard deviation of 3.04 years. Further, comparative analysis showed that the positive rate of absorption elution is more than that of absorption inhibition. Therefore, absorption elution method remains an important tool for determination of blood group of an individual from saliva and it is an aid in forensic identification and in solving medico-legal cases.

Alternative platform for COVID-19 diagnosis based on AuNP-modified lab-on-paper.

COVID-19 has caused global health problems, and so rapid diagnosis is crucial to slow spread of the disease. Herein, a novel lab-on-paper screening method for SARS-CoV-2 Omicron BA.2 variant was developed using a gold nanoparticle-based colorimetric biosensor along with sensitive detection of SARS-CoV-2 antigen using laser desorption ionization-mass spectrometry (LDI-MS). As a result of antigen-antibody interaction, in the presence of SARS-CoV-2 antigen the gold nanoparticles undergo aggregation and change color from red to light purple, allowing for rapid determination of SARS-CoV-2 antigen with the naked eye. Furthermore, the lab-on-paper method can be directly applied as a substrate for sensitive quantitation of SARS-CoV-2 antigen in saliva using LDI-MS without the use of a conventional organic matrix and sample preparation. LDI-MS offers early diagnosis with high sensitivity, rapidity without sample preparation and lower cost per test compared with reverse transcriptase-PCR, which is crucial for preventing mortality in patients with underlying conditions. This method showed linearity over 0.01-1 μg mL-1 covering the cut-off value of 0.048 μg mL-1 for COVID-19 detection in human saliva. Moreover, a colorimetric sensor for urea was also fabricated in-parallel, for prediction of COVID-19 severity in patients with chronic kidney disease. The color change upon increasing urea concentration directly reflected kidney damage, which is related to increasing risk of mortality among patients with COVID-19. Hence, this platform might be a potential device for non-invasive diagnosis of SARS-CoV-2 Omicron BA.2 variant, which is the variant of most concern because it is transmitted more rapidly than the original SARS-CoV-2 virus and the Delta variant.

Urine and Saliva: Relevant Specimens for Malaria Diagnosis?

Blood remains the specimen of preference for malaria diagnosis, whether it is for microscopic, nucleic acid-based or biomarker detection of Plasmodium present in a patient. However, concerning the disadvantages of blood drawing, specimens that can be non-invasively collected under non-hygienic settings would come in handy for malaria diagnosis in endemic areas with limited resources. Although the current approaches using saliva or urine might not be as sensitive and specific as using blood, the potential of these two specimens should not be underestimated and efforts in developing diagnostic methods for Plasmodium detection specifically in these two specimens should continue without giving up. This review not only compiles and summarizes the sensitivity and specificity achieved by various detection approaches when using these samples for malaria diagnosis, it also intends to enhance the possibility of using saliva and urine for diagnostic purposes by describing how Plasmodium nucleic acid and antigens may likely be present in these samples. This review may hopefully encourage and motivate researchers in developing saliva- and urine-based diagnostic methods for Plasmodium detection to facilitate the control and eradication of malaria. In summary, the presence of Plasmodium DNA and antigens in urine and saliva makes these two specimens relevant and useful for malaria diagnosis.

Head-to-head comparison of the accuracy of saliva and nasal rapid antigen SARS-CoV-2 self-testing: cross-sectional study

BackgroundThe diagnostic accuracy of unsupervised self-testing with rapid antigen diagnostic tests (Ag-RDTs) is mostly unknown. We studied the diagnostic accuracy of a self-performed SARS-CoV-2 saliva and nasal Ag-RDT in the general population.MethodsThis large cross-sectional study consecutively included unselected individuals aged ge 16 years presenting for SARS-CoV-2 testing at three public health service test sites. Participants underwent molecular test sampling and received two self-tests (the Hangzhou AllTest Biotech saliva self-test and the SD Biosensor nasal self-test by Roche Diagnostics) to perform themselves at home. Diagnostic accuracy of both self-tests was assessed with molecular testing as reference.ResultsOut of 2819 participants, 6.5% had a positive molecular test. Overall sensitivities were 46.7% (39.3–54.2%) for the saliva Ag-RDT and 68.9% (61.6–75.6%) for the nasal Ag-RDT. With a viral load cut-off (≥ 5.2 log10 SARS-CoV-2 E-gene copies/mL) as a proxy of infectiousness, these sensitivities increased to 54.9% (46.4–63.3%) and 83.9% (76.9–89.5%), respectively. For the nasal Ag-RDT, sensitivities were 78.5% (71.1–84.8%) and 22.6% (9.6–41.1%) in those symptomatic and asymptomatic at the time of sampling, which increased to 90.4% (83.8–94.9%) and 38.9% (17.3–64.3%) after applying the viral load cut-off. In those with and without prior SARS-CoV-2 infection, sensitivities were 36.8% (16.3–61.6%) and 72.7% (65.1–79.4%). Specificities were > 99% and > 99%, positive predictive values > 70% and > 90%, and negative predictive values > 95% and > 95%, for the saliva and nasal Ag-RDT, respectively, in most analyses. Most participants considered the self-performing and result interpretation (very) easy for both self-tests.ConclusionsThe Hangzhou AllTest Biotech saliva self Ag-RDT is not reliable for SARS-CoV-2 detection, overall, and in all studied subgroups. The SD Biosensor nasal self Ag-RDT had high sensitivity in individuals with symptoms and in those without prior SARS-CoV-2 infection but low sensitivity in asymptomatic individuals and those with a prior SARS-CoV-2 infection which warrants further investigation.