Intended Use
This microplate-based ELISA (enzyme linked immunosorbent assay) kit is intended for the qualitative detection of Adenovirus antigen in feces. The assay is a useful tool in the diagnosis of active Adenovirus infection in acute or chronic gastroenteritis.
Contents of Kit
1. Anti-Adenovirus Antibody Coated Microplate
2. Anti-Adenovirus Tracer Antibody
3. Tracer Antibody Diluent
4. ELISA Wash Concentrate
5. ELISA HRP Substrate
6. ELISA Stop Solution
7. Adenovirus Antigen Controls
8. Concentrated Patient Sample Diluent
Storage
This test kit must be stored at 2 - 8°C upon receipt. For the expiration date of the kit refer to the label on the kit box. All components are stable until this expiration date.
General Description
Acute diarrheal disease in young children is a major cause of morbidity worldwide and is a leading cause of mortality in developing countries. Research has shown that enteric adenoviruses, primarily Ad40 and Ad41, are a leading cause of diarrhea in many of these children, second only to the rotaviruses. However many different symptoms can manifest, depending on the type of infecting Adenovirus. There are 49 distinct serotypes that can cause infections in humans.
The diarrhea resulting from enteric adenoviruses is longer in duration than that caused by the rotaviruses, usually lasting 7 - 8 days. Adenovirus infections often show up as conjunctivitis, tonsillitis (which may look exactly like strep throat and cannot be distinguished from strep except by throat culture), an ear infection, or croup. Adenoviruses can also cause gastroenteritis (stomach flu). A combination of conjunctivitis and tonsillitis is particularly common with adenovirus infections. Small children are especially prone to develop adenovirus bronchiolitis or pneumonia, both of which can be severe. In babies, adenoviruses can also cause coughing fits that are almost exactly like whooping cough. Adenoviruses can also lead to viral meningitis or encephalitis. Rarely, adenovirus causes inflammation of the urinary bladder (also known as cystitis), producing blood in the urine. In children, adenoviruses may cause acute upper respiratory infections with fever and runny nose. Adenovirus types 1, 2, 3, 5, and 6 are responsible for most of these infections.
Specific diagnosis of the Adenovirus infection is made by identification of the virus in the patient's stool. Enzyme linked immunosorbent assay (ELISA) is the test most widely used to screen clinical specimens.
Citations
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Adenoviruses (AdV) have been used as vectors for various therapeutic purposes since their isolation in the last century, such as gene therapy for cancer and other malignant tumors, vaccine development, and delivery of the CRISPR-Cas9 mechanism. Adenovirus was used as a vector for the SARS-CoV-2 vaccine during the COVID-19 pandemic. AdV can deliver cargo to dividing and non-dividing cells in vitro and in vivo, and its main limiting factor when used in vivo is immune clearance of the cells. AdV transports cargo with a very limited immune response, but is limited by cargo capacity.
The adenovirus genome is encapsidated within an icosahedral capsid, with the major capsid protein hexon forming the icosahedral section. The capsid at the apex consists of penton motifs and fibers. These two proteins play a key role in the early stages of infection, as they are responsible for interacting with cellular receptors. The icosahedral asymmetric unit consists of four hexon trimers, one penton based monomer, one protein III, two proteins VIII and four proteins IX. AdV infecting humans is classified into six species (A-F), which can be further subdivided into 50 serotypes, of which AdV serotype 2 (Ad2) and AdV serotype 5 (Ad5) are most commonly used in the manufacture of recombinant AdV vectors. These vectors usually cause mild respiratory disease and are not carcinogenic. This makes AdV an optimal cargo transport system, and the mild immune response to AdV makes it the most suitable for in vivo gene therapy. AdVs are widely used as replication-defective vectors, i.e., therapeutic transgenes are expressed in replication-defective viral vectors, and tumor cells are transduced with the transgenes, the expression of which produces anticancer effects. The main advantage is that the therapeutic genes are highly expressed in the tumor cells, while the expression of the vector protein is low, which makes the AdV vector safer.
Figure 1. Structure of the AdV capsid
(Source: Gallardo J, et al. 2021)
In order to improve the safety and efficiency of AdV, the AdV genome has been progressively modified from its wild type, from the replicative region of the first generation to the gene region of the third generation, and a reduction in the immune response was able to be observed through the gradual removal of non-essential DNA regions. The modified results showed an increase in AdV gene insertion capacity from about 7kb to 36kb. AdV vectors continue to be improved, including fiber switching, incorporation of ligands in the viral capsid, and hexon modification of the fibers.
Figure 2. AdV genome with specific modifications for increased efficiency
(Source: Syyam A, et al. 2022)
Alternative Names
AdV (Fecal) ELISA
References
1. Syyam A, et al. Adenovirus vector system: construction, history and therapeutic applications. Biotechniques. 2022 Dec;73(6):297-305.
2. Gallardo J, et al. Adenovirus Structure: What Is New? Int J Mol Sci. 2021 May 15;22(10):5240.
The Effect of Habitat Structure Boulder Spacing on Near-Bed Shear Stress and Turbulent Events in a Gravel Bed Channel
WATER
Authors: Golpira, Amir; Huang, Fengbin; Baki, Abul B. M.
Abstract
This study experimentally investigated the effect of boulder spacing and boulder submergence ratio on the near-bed shear stress in a single array of boulders in a gravel bed open channel flume. An acoustic Doppler velocimeter (ADV) was used to measure the instantaneous three-dimensional velocity components. Four methods of estimating near-bed shear stress were compared. The results suggested a significant effect of boulder spacing and boulder submergence ratio on the near-bed shear stress estimations and their spatial distributions. It was found that at unsubmerged condition, the turbulent kinetic energy (TKE) and modified TKE methods can be used interchangeably to estimate the near-bed shear stress. At both submerged and unsubmerged conditions, the Reynolds method performed differently from the other point-methods. Moreover, a quadrant analysis was performed to examine the turbulent events and their contribution to the near-bed Reynolds shear stress with the effect of boulder spacing. Generally, the burst events (ejections and sweeps) were reduced in the presence of boulders. This study may improve the understanding of the effect of the boulder spacing and boulder submergence ratio on the near-bed shear stress estimations of stream restoration practices.
Epidemiology and Outcomes of Early-Onset and Late-Onset Adenovirus Infections in Kidney Transplant Recipients
OPEN FORUM INFECTIOUS DISEASES
Authors: Bruminhent, Jackrapong; Worawichawong, Suchin; Tongsook, Chutatip; Pasomsub, Ekawat; Boongird, Sarinya; Watcharananan, Siriorn P.
Abstract
Objective. Adenovirus (ADV) infection after kidney transplantation (KT) causes significant morbidity. Patient characteristics and outcomes of ADV infection in KT recipients were investigated. Method. All adult KT recipients with ADV infection between January 2015 and June 2019 were included. ADV infection/disease was defined as detection of ADV DNA in clinical specimens/plus symptoms. Clinical and laboratory findings, treatments, and outcomes were assessed. Results. Adenovirus infection was diagnosed in 24 of 751 (3.2%) KT recipients. Twenty (83%) were male with a median age of 47 years (interquartile range [IQR], 36-58). Fifteen (63%) underwent deceased donor KT, and 13 (54%) received induction therapy. Twenty-one (88%) and 4 (17%) patients developed hemorrhagic cystitis and disseminated disease, respectively. There were equal distributions of early-onset (EOI) (<= 3 months) and late-onset (LOI) (>3 months) infections. Patients who were diagnosed with EOI had lower median absolute lymphocyte counts compared with those with LOI (735/mm(3) [IQR, 543-1123] vs 1122/mm(3) [IQR, 784-1344], P = .04). All achieved resolution after reduction of their immunosuppression regimen and 13 (54%) received cidofovir therapy. Eighteen (75%) developed allograft dysfunction, of which 67% were transient. One (4%) underwent nephrectomy for allograft failure and 1 (4%) died (non-ADV-related). Patients with EOI were more likely to receive cidofovir therapy (75% vs 33%, P = .04) and develop other opportunistic infections (75% vs 8%, P < .001). Conclusions. Adenovirus infection after KT typically involves a genitourinary system and transiently impairs an allograft function. Those who developed early infection tend to have more lymphopenia, coinfection, and receive antiviral therapy.
Reagents for Adenovirus Assay Development 
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