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This article, aimed at nonlaboratorians such as healthcare providers, public health professionals, and policymakers, provides basic concepts and terminology to enable better understanding of other manuscripts in this advanced molecular detection journal supplement. This article focuses on 3 aspects of advanced molecular detection: pathogen genomics, bioinformatics, and public health application, while providing additional resources for understanding.

Solid‐state nanopore arrays are emerging as powerful tools for label‐free, ultrasensitive biosensing, yet their implementation has been constrained by inter‐pore crosstalk and limited fabrication uniformity. A multilayer Al2O3/Au/Si3N4 nanopore architecture, produced via helium ion beam lithography, is introduced to address these limitations through structural and materials‐level innovation. Finite‐element analysis identifies a critical inter‐pore spacing approximately 20 times the pore radius as necessary to minimize electric field coupling, enabling rational array design. The membrane structure incorporates a dielectric Al2O3 layer for electrical isolation and an intermediate gold layer for site‐specific aptamer immobilization, confining molecular recognition to the nanopore interior. Arrays with ∼30 nm pores and <5% size variation achieve 300 nm spacing and support statistically independent, parallel signal acquisition. Diverse nanopore arrays with 75 nm pores and 800 nm spacing are utilized for the specific detection of alpha‐fetoprotein. Detection of alpha‐fetoprotein demonstrates label‐free sensing at concentrations down to ∼3 fM across six orders of magnitude in dynamic range. This platform defines a closed‐loop pathway from theoretical modeling to scalable fabrication, establishing a foundation for rational design and high‐throughput deployment of solid‐state nanopore biosensors. The critical pore spacing—approximately twenty times the pore radius—required to minimize electric field coupling is determined via finite element analysis. A multi‐layered Al2O3/Au/Si3N4 nanopore structure fabricated by helium ion beam lithography is proposed, enabling quantitative analysis of the target analyte while mitigating inter‐pore crosstalk.

Chikungunya virus (CHIKV) infection is a re-emerging arboviral disease with no approved vaccine, although numerous options are in development. Before vaccine implementation, disease burden, affected age group, and hospitalization rate information should be documented. In 2019, a sizeable outbreak of the East Central South African genotype of CHIKV occurred in Myanmar, and during this period, a cross-sectional study was conducted in two regions, Mandalay and Yangon, to examine the molecular and seropositivity rate of the CHIKV infection. The participants (1124) included dengue-suspected pediatric patients, blood donors, and healthy volunteers, who were assessed using molecular assays (quantitative real-time RT-PCR), serological tests (anti-CHIKV IgM capture and IgG indirect enzyme-linked immunosorbent assays), and neutralization tests. The tests confirmed the following positivity rates: 11.3% (127/1124) for the molecular assay, 12.4% (139/1124) for the anti-CHIKV IgM Ab, 44.5% (500/1124) for the anti-CHIKV IgG Ab, and 46.3% (520/1124) for the CHIKV neutralizing Ab. The highest rate for the molecular test occurred with the dengue-suspected pediatric patients. The seroprevalence rate through natural infection was higher in the healthy volunteers and blood donors than that in the pediatric patients. The results of this study will help stakeholders determine the criteria for choosing appropriate recipients when a CHIKV vaccine is introduced in Myanmar.

Background Brucellosis is an emerging yet neglected zoonosis that has been reported in Kenya. Epidemiological data on brucellosis in ruminants is readily accessible; however, reports on brucellosis in pigs remain limited. This study sought to detect Brucella infection in pig serum by both serological and molecular techniques. Serum from 700 pigs randomly collected at a centralized abattoir in Nairobi region, Kenya were screened in parallel, using both Rose Bengal Test (RBT) and competitive Enzyme-Linked Immuno-sorbent Assay (cELISA) for antibodies against Brucella spp. All sera positive by RBT and 16 randomly selected negative samples were further tested using conventional PCR targeting bcsp31 gene and real-time PCR (RT-PCR) assays targeting IS711 and bcsp31 genes. Results A prevalence of 0.57% ( n  = 4/700) was estimated using RBT; none of these samples was positive on cELISA. All RBT positive sera were also positive by both PCRs, while two sero-negative samples also tested positive on RT-PCR ( n  = 6/20). Brucella abortus was detected in four out of the six PCR positive samples through a real-time multiplex PCR. Conclusion The detection of antibodies against Brucella spp. and DNA in serum from slaughterhouse pigs confirm the presence of Brucella in pigs. Therefore, investigation of the epidemiology and role of pigs in the transmission of brucellosis in Kenya is needed. Further targeted studies would be useful to systematically quantify and identify the spp. of Brucella in pigs.

Intraguild predation among arthropod predators in agricultural ecosystems may have a negative impact on biological control. At present, there are few direct reports on trophic relationships among participants of predation in field groups. In this study, we measured the feeding choices of Hippodamia variegata (Goeze) towards mummies with different densities of Aphis gossypii Glover. The dynamics of the occurrence of mummies in the cotton field were investigated over 2017–2019. Singleplex PCR and multiplex PCR were used to detect the predation of 2090 H. variegata individuals on aphids and mummies in Xinjiang cotton field, which revealed the intraguild predation frequency between H. variegata and various parasitoids. There was no obvious feeding preference of H. variegata towards live aphids or mummies, which mainly depended on the relative density of prey. Among the four species of aphids detected in H. variegata, A. gossypii had a high detection rate and was the main prey source of the ladybeetle in the cotton filed. Mostly, ladybeetles consumed parasitoids through mummies, with 6.39% directly feeding on adult parasitoids. H. variegata had strong trophic links to both parasitoids and aphids. We established a food web of aphids–primary parasitoids–hyperparasitoids–H. variegata, which can be used to evaluate the pest control ability of H. variegata from a new perspective.

Background: Emergent vector-borne diseases have gained significant attention in recent years due to their increasing prevalence and impact on public health. With its vast geographic and ecological diversity, Algeria has limited available data on the distribution and prevalence of neglected vector-borne diseases. This study aimed to inventory hematopha­gous ectoparasites, including ticks and fleas, collected from domestic and wild animals such as dogs, hedgehogs, cattle, and rodents across diverse biotopes in northwestern Algeria (Mascara, Oran, Tlemcen, Sidi Bel Abbes, Mostaganem, Tiaret, and Ain Temouchent) and southern Algeria (Laghouat). Methods: A total of 984 arthropods, comprising 609 ticks and 375 fleas, were collected from domestic and wild an­i­mals. Among these, 193 ticks and 105 fleas underwent molecular screening for Rickettsia spp. using gltA and ompA gene-specific primers. Results: The minimum infection rate (MIR) for Rickettsia spp. was estimated at 6.37%, assuming one positive individ­ual per pool. Quantitative PCR revealed the presence of Rickettsia massiliae in 1/68 (1.47%) of Rhipicephalus san­guineus ticks and Rickettsia felis in 7/48 (14.58%) of Ctenocephalides felis fleas. Additionally, a novel strain of Rickett­sia sp. was identified in Rhipicephalus sanguineus and Rhipicephalus turanicus. Conclusion: This study expands the understanding of tick- and flea-borne Rickettsia species in Algeria, highlighting the diverse range of ectoparasite-borne pathogens associated with domestic and wild animals. The findings underscore the importance of continued surveillance and molecular characterization to address the public health risks posed by these pathogens.  

This study was aimed to isolation and phenotypic characterization of P. aeruginosa. 75 carp fish samples were collected from fish farms at Mosul city during (September2021 to January 2022). Each sample was placed separately in sterile plastic bags and transferred directly to the microbiology laboratory under cooling conditions. The API20-20E assay was used to confirm P. aeruginosa isolates. Also the isolates were confirmed molecularly by polymerase chain reaction assay using the primer 16srDNA. Through bacterial isolation were obtained (60) isolates, which formed (36.6, 30, 8.3, 25 )% of each of the skin, gills,  intestines and muscles, respectively. The results showed the sensitivity of P. aeruginosa isolates to impenime (IMP10 µg) and cephalosporin (CIP10 µg) reached (86% and 80 %) respectively, followed by levofloxacin (LEV 5 µg) (74%). But resistant (100%) to both amoxicillin (AM10 µg) and tetracycline (TE10 µg). Isolates varied in their sensitivity and resistance to other antibiotics which included, tobramycin (T0B10 µg), gentamycin (CN10 µg) and amikacin (AK30 µg), the study showed that isolates gave positive results for each hemolytic activity, protease, lecithinase and phospholipase virulence factors tests.

Objective: This short study describes the occurrence of pathogenic Leptospira spp. in two major wet markets in Kota Bharu, Kelantan, Malaysia. Materials and Methods: 30 rodents (20 rats and 10 shrews) were caught in 2 wet markets, and a postmortem was performed to extract both kidneys. Molecular diagnosis via polymerase chain reaction (PCR) was conducted to detect leptospiral DNA using universal and pathogenic Leptospira primers, respectively. Results: The results showed that 20/28 (72%) rat samples were detected positive for Leptospira spp, and all shrews were negative. Further sequencing analysis identified L. interrogans and L. borgpetersenii as the most frequently Leptospirosis species from kidney samples. Conclusions: The presented study here sheds light on the presence of pathogenic leptospires har¬boring the rat population in both wet markets in Kelantan, which presents a great public health risk to wet market workers and visitors.

Ducks are the natural reservoir of influenza A virus and the central host for the avian influenza virus (AIV) subtype H5N1, which is highly pathogenic. Semi-scavenging domestic ducks allow for the reemergence of new influenza subtypes which could be transmitted to humans. We collected 844 cloacal swabs from semi-scavenging ducks inhabiting seven migratory bird sanctuaries of Bangladesh for the molecular detection of avian influenza genes. We detected the matrix gene (M gene) using real-time RT–PCR (RT–qPCR). Subtyping of the AIV-positive samples was performed by RT–qPCR specific for H5, H7, and H9 genes. Out of 844 samples, 21 (2.488%) were positive for AIV. Subtyping of AIV positive samples (n = 21) revealed that nine samples (42.85%) were positive for the H9 subtype, five (23.80%) were positive for H5, and seven (33.33%) were negative for the three genes (H5, H7, and H9). We detected the same genes after propagating the virus in embryonated chicken eggs from positive samples. Semi-scavenging ducks could act as carriers of pathogenic AIV, including the less pathogenic H9 subtype. This can enhance the pathogenicity of the virus in ducks by reassortment. The large dataset presented in our study from seven areas should trigger further studies on AIV prevalence and ecology.

This study was designed for the molecular diagnosis and assessment of histopathological changes of ORF infection in sheep and goats in Basra governorate in Iraq. The virus was detected by using -polymerase chain reaction. The samples were taken from the skin of the lips of animals infected with contiguous ecthema. Hundred samples taken from the sheep and 100 samples from goats of suspected animals, the results of the molecular diagnosis showed that 76 (76%) of sheep were infected and 71 (71%) of goats were infected by diagnosing the partial of ORFV037 gene (173bp), and ORFV039 (703bp), the sequences were determined and recorded in the NCBI and drawn the phylogenetic tree. The results of histopathological study showed presented various changes in cutaneous tissues in sheep include hyperplasia of hair follicles, sebaceous gland and sweet gland, parakeratosis, hyalinization of keratin Also to a picture of tumores, inflammation appear clearly in dermis also to hyperplasia of epidermis towered dermis. In goat showed prolifration of epidermis layer and papillae projection, Highly proliferation structures spinosum, sebaceous gland, sweet gland, and proliferation hair follicles with thickening it wall, hyperkeratosis of epidermis, infiltration of inflammatory cells and ulceration with necrotizing of epidermis.