scholarly journals Technical viability of the YF MAC-HD ELISA kit for use in yellow fever-endemic regions

2021 ◽  
Vol 15 (6) ◽  
pp. e0009417
Author(s):  
Christin H. Goodman ◽  
Maurice Demanou ◽  
Mick Mulders ◽  
Jairo Mendez-Rico ◽  
Alison Jane Basile
Keyword(s):  
South America ◽  
Yellow Fever ◽  
Accurate Diagnosis ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Kit ◽  
Wash Buffer ◽  
Antibody Capture ◽  
Vaccination Campaigns ◽  
Arboviral Disease

Yellow fever (YF), an arboviral disease, affects an estimated 200,000 people and causes 30,000 deaths per year and recently has caused major epidemics in Africa and South America. Timely and accurate diagnosis of YF is critical for managing outbreaks and implementing vaccination campaigns. A YF immunoglobulin M (IgM) antibody-capture (MAC) enzyme-linked immunosorbent assay (ELISA) kit, the YF MAC-HD, was successfully introduced starting in 2018 to laboratories in Africa and South America. The YF MAC-HD kit can be performed in 3.5 hours, test up to 24 samples, and includes all reagents necessary to perform the test, except for water used to dilute wash buffer. In 2018 and 2019, a total of 56 laboratory personnel from 39 countries in Africa and South America were trained to use the kit during workshops, followed by take-home YF IgM proficiency testing (PT) exercises. Participants received either a 10- or 20-sample YF PT panel and performed testing using the YF MAC-HD kit. All countries obtained 90% or higher correct results. These results verified the technical viability and transferability of YF MAC-HD kit use for laboratories in YF-endemic countries.

West Nile Virus

2016 ◽  
Author(s):  
Matthew Finn
Keyword(s):  
West Nile Virus ◽  
Rna Virus ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mosquito Vector ◽  
West Nile ◽  
Csf Analysis ◽  
Neuroinvasive Disease ◽  
Breeding Grounds ◽  
Arboviral Disease

West Nile virus (WNV) is a single-stranded RNA virus of the Flavivirus family that is transmitted via a mosquito vector, typically causing fever and capable of causing meningoencephalitis. Although mortality is low, it can lead to debilitating neuroinvasive disease in some patients. WNV is a leading cause of domestically-acquired arboviral disease and most commonly occurs in late August and early September. Consider WNV in otherwise unexplained cases of meningitis or encephalitis. Initial testing should consist of cerebrospinal fluid (CSF) analysis and West Nile immunoglobulin M enzyme-linked immunosorbent assay in serum and/or CSF. WNV is a nationally notifiable disease. Prevention remains the key to controlling this disease. Reducing the breeding grounds of the Culex mosquito and using insect repellant to prevent bites are two important strategies.

scholarly journals Standardization of Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assays for Routine Diagnosis of Arboviral Infections

2000 ◽  
Vol 38 (5) ◽  
pp. 1823-1826 ◽  
Author(s):  
Denise A. Martin ◽  
David A. Muth ◽  
Teresa Brown ◽  
Alison J. Johnson ◽  
Nick Karabatsos ◽  
...  
Keyword(s):  
Immunoglobulin M ◽  
Detection Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Virus Family ◽  
Serum Dilution ◽  
Routine Diagnosis ◽  
Human Sera ◽  
Antiviral Antibody ◽  
Antibody Capture

Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) is a rapid and versatile diagnostic method that readily permits the combination of multiple assays. Test consolidation is especially important for arthropod-borne viruses (arboviruses) which belong to at least three virus families: the Togaviridae,Flaviviridae, and Bunyaviridae. Using prototype viruses from each of these families and a panel of well-characterized human sera, we have evaluated and standardized a combined MAC-ELISA capable of identifying virus infections caused by members of each virus family. Furthermore, by grouping antigens geographically and utilizing known serological cross-reactivities, we have reduced the number of antigens necessary for testing, while maintaining adequate detection sensitivity. We have determined that a 1:400 serum dilution is most appropriate for screening antiviral antibody, using a positive-to-negative ratio of ≥2.0 as a positive cutoff value. With a blind-coded human serum panel, this combined MAC-ELISA was shown to have test sensitivity and specificity that correlated well with those of other serological techniques.

scholarly journals Development of a Human-Murine Chimeric Immunoglobulin M Antibody for Use in the Serological Detection of Human Flavivirus Antibodies

2009 ◽  
Vol 16 (5) ◽  
pp. 679-685 ◽  
Author(s):  
Brett A. Thibodeaux ◽  
John T. Roehrig
Keyword(s):  
Yellow Fever Virus ◽  
Positive Control ◽  
Encephalitis Virus ◽  
Immunoglobulin M ◽  
Chimeric Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Variable Regions ◽  
Igm Antibody ◽  
Antibody Capture ◽  
Serological Activity

ABSTRACT Current diagnosis of human flaviviral infections relies heavily on serological techniques such as the immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA). Broad application of this assay is hindered by a lack of standardized human positive-control sera that react with the wide variety of flaviviruses that can cause human disease, e.g., dengue virus (DENV), West Nile virus (WNV), yellow fever virus (YFV), Japanese encephalitis virus (JEV), and St. Louis encephalitis virus (SLEV). We have created a human-murine chimeric antibody combining the variable regions of the broadly flavivirus cross-reactive murine monoclonal antibody (MAb) 6B6C-1 and the constant region of human IgM to produce a standardized reagent capable of replacing human positive-control sera in a MAC-ELISA for the diagnosis of all human flaviviral infections. The human-murine chimeric IgM antibody secreted from plasmid-transformed Sp2/0-Ag14 cells had a level of serological activity identical to that of 6B6C-1 as measured by ELISA, immunoblotting, and MAC-ELISA for multiple members of the flavivirus genus, including WNV, SLEV, YFV, DENV, and JEV.

scholarly journals Serotype-Cross-Reactive Immunoglobulin M Responses in Dengue Virus Infections Determined by Enzyme-Linked Immunosorbent Assay

2000 ◽  
Vol 7 (5) ◽  
pp. 774-777 ◽  
Author(s):  
Masaru Nawa ◽  
Ken-Ichiro Yamada ◽  
Tomohiko Takasaki ◽  
Toshitaka Akatsuka ◽  
Ichiro Kurane
Keyword(s):  
Dengue Virus ◽  
Japanese Patients ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Serum Samples ◽  
Reverse Transcription Pcr ◽  
Virus Serotype ◽  
Dengue Virus Serotypes ◽  
Antibody Capture

ABSTRACT We developed immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assays (ELISAs) with four monovalent dengue virus antigens. We attempted to determine whether IgM responses in dengue virus infections are serotype specific or serotype cross-reactive. Serum samples from 14 confirmed dengue cases were examined. In these 14 cases, which consisted of 12 Japanese and 2 non-Japanese patients, infecting dengue virus serotypes were defined by reverse transcription-PCR. Thirteen of the 14 cases were IgM positive in ELISA. IgM responses were serotype cross-reactive in these 13 cases but were highest against infecting dengue virus serotype in 9 of the 13 cases. These results indicate that IgM responses are generally dengue serotype cross-reactive but that IgM levels are highest against the infecting serotype in most dengue cases.

Viral Causes of Acute Febrile Jaundice in Selected Provinces of Zambia

2021 ◽  
Author(s):  
Boniface Kabungo ◽  
AaroAyato Takada ◽  
Masahiro Kajihara ◽  
Akina Mori ◽  
Katendi Changula ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Fever ◽  
Yellow Fever ◽  
Hepatitis A ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Serum Samples ◽  
Polymerase Chain ◽  
Febrile Jaundice

Abstract Background Following the yellow fever (YF) risk assessment conducted in 2013, Ministry of Health in collaboration with WHO successfully implemented YF case based surveillance among the YF suspects in the high risk areas of Zambia. To date, none of the patients has been confirmed as a case of YF and the epidemiology of flavi-viruses has not been comprehensively investigated in Zambia. As YF may be hardly distinguished clinically from other febrile diseases, because early in the clinical course YF may appear similar to other diseases but YF will diverge clinically as the disease course progresses. This study was designed to investigate the viral causes of febrile jaundice among YF suspects in selected provinces of Zambia. Method We conducted a retrospective study on 93 archived serum samples previously collected from patients meeting a case definition of YF suspect from January 2014 to July 2015 presented in selected health facilities. Yellow Fever, Dengue Fever, West Nile, pan Flavivirus, and Hepatitis A viruses were tested by reverse transcriptase polymerase chain reaction (RT_PCR) and Hepatitis B virus using PCR, while Hepatitis C and Hepatitis E viruses were tested by nested polymerase chain reaction (nPCR). Samples were also tested for YF and dengue fever (DF) antibodies using in-house immunoglobulin M enzyme linked immunosorbent assay (Ig M ELISA) and immunoglobulin M rapid test respectively. STATA version 12 was used for data analysis. Results Fourteen percent (13/93) of the serum samples were identified as YF IgM positive. None of the samples tested positive for DF IgM ELISA. All 93 serum samples tested negative for the flaviviruses by RT-PCR. However, 8.6% (8/93) showed acute Hepatitis A and 2/20 (10%) of pooled sera tested positive for HBV. The median age of patients with Hepatitis A was 9.5 years old and for those without evidence of HAV infection was 19 years old. Approximately 85 (91.4%) of patients had acute diseases of unknown origin. Conclusion The study revealed that YF IgM was prevalent among study participants. However, the causes of fever and jaundice in Zambia may include viral hepatitis and needs to be considered if flaviviral diseases are suspected.

scholarly journals Recombinant Multiepitope Protein for Diagnosis of Leptospirosis

2008 ◽  
Vol 15 (11) ◽  
pp. 1711-1714 ◽  
Author(s):  
Xu'ai Lin ◽  
Yin Chen ◽  
Jie Yan
Keyword(s):  
Outer Membrane Proteins ◽  
Agglutination Test ◽  
Accurate Diagnosis ◽  
Immunoglobulin M ◽  
Severe Illness ◽  
Enzyme Linked Immunosorbent Assay ◽  
Promising Alternative ◽  
Immunity Reaction ◽  
Gene Recombinant ◽  
Immunodominant Epitopes

ABSTRACT Leptospirosis is an emerging infectious disease and is considered to be the most widespread zoonotic disease in the world. It can be misdiagnosed because manifestations of this febrile disease vary from mild flu-like symptoms to severe illness involving vital organs such as the liver and lungs. Therefore, accurate diagnosis for differentiation of leptospirosis from other pyrogenic infections prevailing in the same locality is imperative for proper treatment. Here, we report a customized recombinant leptospirosis multiepitope protein (r-LMP) that can specifically detect the immunoglobulin class of anti-leptospirosis antibodies in patient sera. Immunodominant epitopes from leptospire outer membrane proteins OmpL1, LipL21, and LipL32 were predicted and confirmed using phage display and immunity reaction. On the basis of the sequences of the identified epitopes, five major immunodominant epitopes were selected to construct a synthetic gene, recombinant lmp. The recombinant lmp gene was doubled and expressed in Escherichia coli. The recombinant protein was purified and used as an antigen to develop an enzyme-linked immunosorbent assay for detection of special immunoglobulin M (IgM) or IgG in sera from patients with leptospirosis or other febrile illnesses and healthy subjects. The results showed that the r-LMP protein recognized IgG and IgM in all the sera that were microscope agglutination test positive, and there were no cross-reactions with other patient sera. This approach of creating customized antigens coupled to overexpression and simple purification offers a promising alternative option for leptospirosis diagnosis, with the potential to circumvent the drawbacks of whole-leptospirosis-antigen-based assays.

scholarly journals Development of a Human-Murine Chimeric Immunoglobulin M for Use in the Serological Detection of Human Alphavirus Antibodies

2011 ◽  
Vol 18 (12) ◽  
pp. 2181-2182 ◽  
Author(s):  
Brett A. Thibodeaux ◽  
Nathan M. Liss ◽  
Amanda N. Panella ◽  
John T. Roehrig
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Immunoglobulin M ◽  
Murine Monoclonal Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Detection ◽  
Antibody Capture

ABSTRACTDiagnosis of human alphaviral infections relies on serological techniques, such as the immunoglobulin M antibody capture–enzyme-linked immunosorbent assay (MAC-ELISA). We have humanized the alphavirus broadly cross-reactive murine monoclonal antibody 1A4B-6 to create a reagent capable of replacing human positive sera in the MAC-ELISA for diagnosis of human alphaviral infections.

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