scholarly journals Performance Evaluation of Commercial Dengue Diagnostic Tests for Early Detection of Dengue in Clinical Samples

2017 ◽  
Vol 2017 ◽  
pp. 1-4 ◽  
Author(s):  
Tuan Nur Akmalina Mat Jusoh ◽  
Rafidah Hanim Shueb
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Dengue Infection ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Kappa Agreement

The shattering rise in dengue virus infections globally has created a need for an accurate and validated rapid diagnostic test for this virus. Rapid diagnostic test (RDT) and reverse transcription-polymerase chain reaction (RT-PCR) diagnostic detection are useful tools for diagnosis of early dengue infection. We prospectively evaluated the diagnostic performance of nonstructural 1 (NS1) RDT and real-time RT-PCR diagnostic kits in 86 patient serum samples. Thirty-six samples were positive for dengue NS1 antigen while the remaining 50 were negative when tested with enzyme-linked immunosorbent assay (ELISA). Commercially available RDTs for NS1 detection, RTK ProDetect™, and SD Bioline showed high sensitivity of 94% and 89%, respectively, compared with ELISA. GenoAmp® Trioplex Real-Time RT-PCR and RealStar® Dengue RT-PCR tests presented a comparable kappa agreement with 0.722. The result obtained from GenoAmp® Real-Time RT-PCR Dengue test showed that 14 samples harbored dengue virus type 1 (DENV-1), 8 samples harbored DENV-2, 2 samples harbored DENV-3, and 1 sample harbored DENV-4. 1 sample had a double infection with DENV-1 and DENV-2. The NS1 RDTs and real-time RT-PCR tests were found to be a useful diagnostic for early and rapid diagnosis of acute dengue and an excellent surveillance tool in our battle against dengue.

scholarly journals Real-time PCR for detecting circulating dengue virus in the Guangdong Province of China in 2006

2008 ◽  
Vol 57 (12) ◽  
pp. 1547-1552 ◽  
Author(s):  
Zhijun Bai ◽  
Licheng Liu ◽  
Zeng Tu ◽  
Lisi Yao ◽  
Jianwei Liu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Cross Reactivity ◽  
Clinical Samples ◽  
Sequencing Analysis ◽  
Serum Samples ◽  
Serological Tests ◽  
Wide Range ◽  
Pcr Assays

Dengue virus (DENV) causes a wide range of diseases in humans, from the acute febrile illness dengue fever (DF) to life-threatening dengue haemorrhagic fever/dengue shock syndrome. We developed four real-time quantitative PCR assays for each serotype of DENV based on computational analysis. These assays had high sensitivity and specificity without cross-reactivity for the four serotypes. To evaluate the performance of these assays in detecting and typing the virus in clinical samples, we analysed 64 serum samples from Guangdong during 2006. The results showed that 71 % of those samples were positive by the DEN-1 assay. The DENV assay results, in agreement with the serological tests and sequencing analysis, showed that the pathogen resulting in the DF explosion in Guangdong in 2006 belonged to DEN-1. Compared to the serological assays, the real-time PCR assays that we developed were much more sensitive in the 1–3 days after onset of the symptoms.

scholarly journals Co-Circulation of Dengue Serotypes in Coastal Andhra Pradesh, India - A Descriptive Study

2020 ◽  
Vol 9 (40) ◽  
pp. 2965-2969
Author(s):  
Suryamani Chintapalli ◽  
Apparao Peddepalli ◽  
Sivajyothi Pilli ◽  
Monika Deepthi Pilli ◽  
Kanaka Mahalakshmi Yandra
Keyword(s):  
Dengue Virus ◽  
Andhra Pradesh ◽  
Febrile Illness ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Medical College ◽  
Igm Antibodies ◽  
Dengue Outbreaks ◽  
Ns1 Antigen

BACKGROUND Dengue is an acute febrile illness caused by mosquito-borne dengue viruses (DENV S) consisting of four serotypes (DENV 1 - 4) from flaviviridae family, genus flavivirus. These four are antigenically related serotypes designated as DEN V - 1, DEN V - 2, DEN V - 3 and DEN V – 4. In this context, the present study focuses on the circulating serotypes of dengue in coastal Andhra Pradesh. METHODS Study was done at Andhra Medical College, Visakhapatnam, teaching hospital in Andhra Pradesh. Acute phase dengue serum samples were collected and tested for NS1 antigen and antihuman IgM antibodies by enzyme linked –immunosorbent assay (ELISA). NS1 positive samples were further serotyped by reverse transcriptase real time polymerase chain reaction (R RT - PCR). RESULTS A total of 796 serum samples were included in the study. 300 (37.7 % ) samples were positive for NS1 and IgM antibodies. 192 NS1 antigen positive samples were further processed for serotyping by r RT PCR. Among these samples 72 were negative by r RT PCR. DENV-2 (41 %) was the predominant serotype followed by DENV-4 (37 %), DENV-3 (12 %) and DENV-1 (10 %) in the descending order. CONCLUSIONS All the four dengue serotypes are in co-circulation. Among all the four types, DENV-2 was predominant, followed by DENV-4. By knowing the predominant serotype in circulation, we can forecast dengue outbreaks and take necessary measures like control of vectors. KEY WORDS Andhra Pradesh, Dengue Virus, Dengue Virus - 2, Dengue Virus - 4, Outbreak, Serotypes

scholarly journals Potential Point-of-Care Testing for Dengue Virus in the Field

2018 ◽  
Vol 56 (5) ◽  
Author(s):  
Wei-Kung Wang ◽  
Duane J. Gubler
Keyword(s):  
Dengue Virus ◽  
Diagnostic Test ◽  
Reverse Transcription ◽  
Point Of Care ◽  
Detection Methods ◽  
Clinical Samples ◽  
Complete Agreement ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Denv Serotypes

ABSTRACT The four serotypes of dengue virus (DENV) cause one of the most important and rapidly emerging mosquito-borne viral diseases in humans. Of the currently available diagnostic tests for dengue, the reverse transcription-PCR (RT-PCR) assay is the most sensitive and specific, and so it is commonly used as the gold standard. However, the requirement of a sophisticated and expensive thermal cycler makes it very difficult to use as a point-of-care diagnostic test in resource-limited regions where dengue is endemic. Tsai et al. (J Clin Microbiol 56:e01865-17, 2018, https://doi.org/10.1128/JCM.01865-17 ) report the analytical and clinical performances of a reverse transcription-insulated isothermal PCR (RT-iiPCR) assay with a portable nucleic acid analyzer for rapid detection of the four DENV serotypes; its reproducibility and complete agreement on clinical samples with the multiplex RT-PCR assay developed by the Centers for Disease Control and Prevention suggest that the dengue RT-iiPCR is a potential point-of-care test. Compared with other DENV RNA detection methods, the unique isothermal PCR design of RT-iiPCR, together with further improvements, would represent a promising new type of field-deployable diagnostic test for dengue.

scholarly journals Detection of anti-HEV antibodies and RNA of HEV in pigs from a hyperendemic Italian region with high human seroprevalence

2020 ◽  
Author(s):  
Camillo Martino ◽  
Elisa Rampacci ◽  
Ilaria Pierini ◽  
Monica Giammarioli ◽  
Valentina Stefanetti ◽  
...  
Keyword(s):  
Real Time ◽  
Central Italy ◽  
Meat Products ◽  
Human Infection ◽  
Zoonotic Transmission ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Viral Origin ◽  
South Central

Abstract Background Pigs are considered the main reservoir of genotypes 3 and 4 of hepatitis E virus (HEV), which is the major cause of acute hepatitis of viral origin in humans worldwide. An increasing number of autochthonous HEV infections have been observed in recent years in industrialized countries, most likely as a result of zoonotic transmission through the consumption of raw or undercooked meat products. Methods Two hundred and thirty-three blood and liver samples were collected at four different local slaughterhouses from domestic pigs bred in Abruzzo, a region of south-central Italy, where there is the highest human seroprevalence to HEV compared with the rest of Italy. An indirect enzyme-linked immunosorbent assay kit was used for detecting anti-HEV IgG in the sera, while the presence of HEV RNA was investigated by performing a real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Results Between 87.3% and 100% of swine serum samples collected in different slaughterhouses of Abruzzo were positive for anti-HEV antibodies. Conversely, none of the liver samples collected from the same animals were positive for HEV by real-time RT-PCR. Conclusions The hypothesis of foodborne zoonotic transmission from local pigs as responsible for the hyperendemic status of Abruzzo cannot be corroborated. However, the high seroprevalence observed in pigs indicates that HEV is highly circulating in these territories. We propose to further investigate the role of wild fauna and trade in carrier pigs, and the maintenance of HEV virulence in the environment and meat supply chain to shed light on the possible sources of human infection and the degree of occupational risk.

scholarly journals One-step real-time RT-PCR assays for serotyping dengue virus in clinical samples

2015 ◽  
Vol 15 (1) ◽  
Author(s):  
Erik Alm ◽  
Gunnel Lindegren ◽  
Kerstin Ingrid Falk ◽  
Nina Lagerqvist
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Clinical Samples ◽  
Rt Pcr ◽  
One Step ◽  
Pcr Assays

Detection of a novel bunyavirus associated with fever, thrombocytopenia and leukopenia syndrome in Henan Province, China, using real-time reverse transcription PCR

2013 ◽  
Vol 62 (7) ◽  
pp. 1060-1064 ◽  
Author(s):  
Xueyong Huang ◽  
Licheng Liu ◽  
Yanhua Du ◽  
Hongxia Ma ◽  
Yujiao Mu ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Henan Province ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Serum Samples ◽  
Reverse Transcription Pcr

A novel bunyavirus associated with fever, thrombocytopenia and leukopenia syndrome (FTLS) was discovered in Henan Province, China. Here, we report the development of an assay for this novel bunyavirus based on real-time reverse transcription PCR (RT-PCR). The assay exhibited high sensitivity and specificity without cross-reactivity towards 13 other viruses that cause similar symptoms. To evaluate the performance of this assay in detecting clinical samples, we analysed 261 serum samples from patients in Henan Province between 2007 and 2010. Of these samples, 91.95 % were bunyavirus positive. Compared with serological assays, the real-time PCR assay was much more sensitive in identifying infected patients 1 to 7 days after the onset of symptoms.

scholarly journals Dengue Virus Detection by Serological and Molecular Method in Different Hospitals of Nepal

2013 ◽  
Vol 11 (2) ◽  
pp. 24-28
Author(s):  
Yogendra Shah ◽  
Govind Prasad Gupta ◽  
Kishor Pandey ◽  
Sher Bahadur Pun ◽  
Krishna Prasad Pant ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Age Groups ◽  
Virus Detection ◽  
Public Health Problem ◽  
Viral Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Age Group ◽  
Rt Pcr ◽  
Serum Samples ◽  
Disease Hospital

Introduction: Dengue is an emerging mosquito-borne viral disease in the world and is the serious public health problem of Nepal. Methods: This study was designed to determine sero-epidemiology of dengue virus infection during the period (June-Nov) of 2010 among suspected patients with fever visiting Koshi Zonal Hospital (KZH), Biratnagar, Narayani sub-regional Hospital (NSH), Birgunj, Sukraraj Tropical and Infectious Disease Hospital (STIDH), Kathmandu and Dhading District Hospital (DDH), Dhadingbeshi. The sero-prevalence of anti-dengue IgM antibody was determined by enzyme linked immunosorbent assay (ELISA). Results: Among 271 serum samples tested, the anti-dengue IgM positivity was 14.4%. Sero-positivity in male was 10.7% of total and that in female was 3.7%.  Among different age groups, the highest positive cases 11.8% were from age group 15-50 years and found least among the age group above 50 years 0.4%. Out of 4 different hospitals, the highest positive positive cases from STIDH with 9.2% and the least positive cases were from DDH (0.4%). RT-PCR showed 4.7% positivity of 21 samples tested. Conclusions: Enzyme immunoassay and RT-PCR serological marker can be used to diagnose the acute patients of dengue during outbreaks.Medical Journal of Shree Birendra Hospital; July-December 2012/vol.11/Issue2/24-27 DOI: http://dx.doi.org/10.3126/mjsbh.v11i2.7905 

scholarly journals Investigations, actions and learning from an outbreak of SARS-CoV-2 infection among healthcare workers in the United Kingdom

2020 ◽  
pp. 175717742097679
Author(s):  
Kordo Saeed ◽  
Emanuela Pelosi ◽  
Nitin Mahobia ◽  
Nicola White ◽  
Christopher Labdon ◽  
...  
Keyword(s):  
Real Time ◽  
Healthcare Workers ◽  
Healthcare Facility ◽  
Rt Pcr ◽  
Serum Samples ◽  
The United Kingdom ◽  
Key Issues ◽  
Current Infection ◽  
Polymerase Chain ◽  
A Current

Background: We report an outbreak of SARS coronavirus-2 (SARS-CoV-2) infection among healthcare workers (HCW) in an NHS elective healthcare facility. Methodology: A narrative chronological account of events after declaring an outbreak of SARS-CoV-2 among HCWs. As part of the investigations, HCWs were offered testing during the outbreak. These were: (1) screening by real-time reverse transcriptase polymerase chain reaction (RT- PCR) to detect a current infection; and (2) serum samples to determine seroprevalence. Results: Over 180 HCWs were tested by real-time RT-PCR for SARS-CoV-2 infection. The rate of infection was 15.2% (23.7% for clinical or directly patient-facing HCWs vs. 4.8% in non-clinical non-patient-facing HCWs). Of the infected HCWs, 57% were asymptomatic. Seroprevalence (SARS-CoV-2 IgG) among HCWs was 13%. It was challenging to establish an exact source for the outbreak. The importance of education, training, social distancing and infection prevention practices were emphasised. Additionally, avoidance of unnecessary transfer of patients and minimising cross-site working for staff and early escalation were highlighted. Establishing mass and regular screening for HCWs are also crucial to enabling the best care for patients while maintaining the wellbeing of staff. Conclusion: To our knowledge, this is the first UK outbreak report among HCWs and we hope to have highlighted some key issues and learnings that can be considered by other NHS staff and HCWs globally when dealing with such a task in future.

scholarly journals Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
Author(s):  
Manphool S. Fageria ◽  
Mathuresh Singh ◽  
Upeksha Nanayakkara ◽  
Yvan Pelletier ◽  
Xianzhou Nie ◽  
...  
Keyword(s):  
Real Time ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Current Season ◽  
Seed Market ◽  
Polymerase Chain ◽  
Time Of Harvest

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

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