Effectiveness of Reverse Transcription-PCR, Virus Isolation, and Enzyme-Linked Immunosorbent Assay for Diagnosis of Influenza A Virus Infection in Different Age Groups

Steininger, Christoph; Kundi, Michael; Aberle, Stephan W.; Aberle, Judith H.; Popow‐Kraupp, Theresia

doi:10.1128/jcm.40.6.2051-2056.2002

Cited by 122 publications

(115 citation statements)

References 22 publications

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“…For rhinovirus, the aliquots of RNA samples were reversed in two cDNA according to the method of Steininger et al (17). For the detection of influenza A virus, we used the methods of Steininger et al (18). For the influenza B virus, a rapid single-step RT-PCR assay was carried out according to the method of Daum et al (19).…”

Section: Methodsmentioning

confidence: 99%

Soluble Adhesion Molecules and Cytokines in Children Affected by Recurrent Infections of the Upper Respiratory Tract

et al. 2004

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The objective of this study was to compare plasma levels of soluble adhesion molecules and Th1-Th2 type cytokines in 44 children with frequently recurrent respiratory infections (FRRI) of upper airways, defined as having nine or more episodes per year, and in 34 children without recurrence; all subjects were followed-up for 12 mo. The viral etiology was determined by cultures from nasal, pharyngeal, and ear secretions, using PCR and immunofluorescence. Plasma levels of five soluble adhesion molecules (E-selectin, P-selectin, L-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1) and interferon (IFN)-␥, IL-12, IL-4, and IL-10 were measured in patients and in 15 healthy controls using sandwich ELISA. During acute phase, all patients showed significant increase in plasma levels of soluble adhesion molecules; during the followup, the levels were greater in children with FRRI. A difference of cytokine profile was demonstrated between the patients with and without FRRI: an increased IL-4 and IL-10 release with decreased levels of IFN-␥ and IL-12 suggested a skewing into Th2-type response, in patients with FRRI. This pattern persisted during the follow-up. In patients without recurrence, an increased IFN-␥ and IL-12 release, together with decreased levels of IL-4 and IL-10, showed a skewing into Th1-type responses; in the follow-up these cytokines reached normal values. In conclusion, the abnormal levels of all examined parameters in children with FRRI may reflect the persistence of an inflammatory microenvironment in the airways and an activation of the immune system that may contribute to the frequently recurrence of the respiratory disease. Infections of the URTI are the most common cause of viral morbidity in children. They represent the most frequent managed problem in general pediatric practice (1) and are responsible for more than one third of school absences (2). Many different immunologic problems can concur to the development of respiratory infections (3). Disorders of the aspecific humoral and the specific cellular defense are the principal cause of inflammation in the upper respiratory diseases (4). The recruitment and activation of inflammatory cells at the site of infection involve the expression of leukocyte and vascular adhesion molecules and the generation of pro-inflammatory cytokines, which contribute to the injury of lung epithelial and endothelial barriers (5). An important role in the early transient adhesion phases of leukocytes is mediated by the selectin family, which includes three distinct carbohydrate receptors expressed by either endothelial cells (E-selectin), leukocytes (L-selectin), or platelets and endothelial cells (P-selectin) (6). ICAM-1 and VCAM-1 are membrane glycoproteins, belonging to the immunoglobulin superfamily; ICAM-1 are widely expressed on respiratory epithelial cells, monocytes, macrophages, dendritic cells, and, together with VCAM-1, on vascular endothelial cells (7,8). These adhesion molecules, after a proteolytic cleavage, are found as so...

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Section: Methodsmentioning

confidence: 99%

Soluble Adhesion Molecules and Cytokines in Children Affected by Recurrent Infections of the Upper Respiratory Tract

et al. 2004

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“…According to Steininger et al (2002), the most suitable tests for effectively diagnosing avian viral diseases, including avian Influenza, WNV and NDV, are RT-PCR and qRT-PCR owing to their high sensitivity and specificity. Based on these data, we analyzed the samples from the captured birds for the three viruses, using molecular methods.…”

Section: Discussionmentioning

confidence: 99%

Investigation of Influenza A, West Nile and Newcastle Disease Viruses in Birds from the Pantanal Wetlands of Mato Grosso, Brazil

Pinto

Ometto

Araújo

et al. 2016

Rev. Bras. Cienc. Avic.

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The Pantanal is the world's largest wetland biome with a seasonal flood pulse that attracts a great diversity of birds, many of which are migratory. Birds can be natural reservoirs Influenza A, West Nile and Newcastle Disease viruses. However, the occurrence of carriers for these viruses in the Pantanal was not verified yet. The present study evaluated the occurrence of natural infection by Influenza A, WN and ND virus of birds in the municipality of Poconé, a subregion of the Pantanal in the state of Mato Grosso, Brazil. A total of 76 birds belonging to 11 orders and 20 families were captured using mist nets. The most representative order was Passeriformes, followed by the other nine orders, which included Columbiformes, Psittaciformes, Charadriiformes and Anseriformes. The most representative family was Thamnophilidae, with 16 individuals (21.0%), followed by the family Tyrannidae with 10 individuals (7.6%) and the family Furnariidae, with eight individuals (10.5%). The bird species were identified, and cloacal and tracheal swab samples were collected. The samples were subjected to RNA extraction and tested for the presence of the three agents by real-time polymerase chain reaction (qRT-PCR). All the sampled birds were considered healthy, had no clinical sign of infection, and were tested negative for the three viruses. Based on our findings, we can conclude that Influenza, West Nile and Newcastle Disease viruses were absent from the samples in this region of the Pantanal wetlands during the period of this study.

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“…Notwithstanding, only the use of expensive transport kits is not a guarantee of yielding precise diagnostic results. All specimens must be stored refrigerated (recommended between 2º to 8 ºC) for up to 24 hours prior the tests without drastically affecting test performance (Steininger et al 2002). It is emphasized that samples under long storage and constant freezing and thawing will possibly degrade viral RNA and cDNA.…”

Section: The Importance Of the Collected Samplesmentioning

confidence: 99%

Diagnostic Methods of Influenza a From Clinical Specimens: A Critical Review of Literature

Bello¹,

Lima

Azevedo³

2015

VR&R

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Effectiveness of Reverse Transcription-PCR, Virus Isolation, and Enzyme-Linked Immunosorbent Assay for Diagnosis of Influenza A Virus Infection in Different Age Groups

Cited by 122 publications

References 22 publications

Soluble Adhesion Molecules and Cytokines in Children Affected by Recurrent Infections of the Upper Respiratory Tract

Soluble Adhesion Molecules and Cytokines in Children Affected by Recurrent Infections of the Upper Respiratory Tract

Investigation of Influenza A, West Nile and Newcastle Disease Viruses in Birds from the Pantanal Wetlands of Mato Grosso, Brazil

Diagnostic Methods of Influenza a From Clinical Specimens: A Critical Review of Literature

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