Abstract
Introduction
Correct diagnosis of Lyme disease (LD) still represents a big challenge worldwide. LD is caused by spirochetes of the Borrelia burgdorferi sensu lato (Bbsl) complex and is transmitted to humans by the bite of infected ticks. In North America, Borrelia burgdorferi sensu stricto (Bbss) is the dominant genospecies associated with human LD; in Eurasia B. afzelii and B. garinii, are the two most prevalent pathogenic species. Scientific evidence suggests that different Bbsl genospecies differ in terms of expressed antigens, disease presentations, response to antibiotics and different clinical manifestations have also been recognized due to a different organotropism. This introduces diagnostic issues and poses additional question marks about the optimal management of patients. Here, we describe a multiplex peptide ELISA to allow serological genospecies identification of the three key Bbsl species.
Materials and Methods
Diagnostic peptide epitopes were chosen through (a) systematic review of outer surface proteins (Osp) of immune-diagnostic value, (b) selection of proteins with evidence of ortholog sequence divergence and (c), B cell epitope prediction and modelling. Genospecies ortholog specific epitopes from 7 oligopeptide regions in 5 Osps were finally synthesised and evaluated in a multiplex peptide ELISA testing a panel of US and EU early-stage and late-stage Lyme disease sera.
Results
The 7 oligopeptide panel achieved the desired sensitivity and accuracy. Sensitivity and correct genospecies identification were 56.2% and 88.8% respectively for IgM responses, and 71.8% and 95.6% respectively for IgG, responses. Higher diagnostic power was observed when IgM and IgG antibody detection was combined. This gave sensitivity and correct genospecies identification of 90.6% and 96.5%, respectively.
Discussion
The multiplex ELISA platform is highly effective in discrimination of infections with Bbss, B. garinii or B. afzelii. In the longer term, diagnostic peptides could be incorporated into a point of care rapid test.
Correct diagnosis of Lyme disease (LD) still represents a big challenge worldwide. LD is caused by spirochetes of the Borrelia burgdorferi sensu lato (Bbsl) complex and is transmitted to humans by the bite of infected ticks. In North America, Borrelia burgdorferi sensu stricto (Bbss) is the dominant genospecies associated with human LD; in Eurasia B. afzelii and B. garinii, are the two most prevalent pathogenic species. Scientific evidence suggests that different Bbsl genospecies differ in terms of expressed antigens, disease presentations, response to antibiotics and different clinical manifestations have also been recognized due to a different organotropism. This introduces diagnostic issues and poses additional question marks about the optimal management of patients. Here, we describe a multiplex peptide ELISA to allow serological genospecies identification of the three key Bbsl species.
Materials and Methods
Diagnostic peptide epitopes were chosen through (a) systematic review of outer surface proteins (Osp) of immune-diagnostic value, (b) selection of proteins with evidence of ortholog sequence divergence and (c), B cell epitope prediction and modelling. Genospecies ortholog specific epitopes from 7 oligopeptide regions in 5 Osps were finally synthesised and evaluated in a multiplex peptide ELISA testing a panel of US and EU early-stage and late-stage Lyme disease sera.
Results
The 7 oligopeptide panel achieved the desired sensitivity and accuracy. Sensitivity and correct genospecies identification were 56.2% and 88.8% respectively for IgM responses, and 71.8% and 95.6% respectively for IgG, responses. Higher diagnostic power was observed when IgM and IgG antibody detection was combined. This gave sensitivity and correct genospecies identification of 90.6% and 96.5%, respectively.
Discussion
The multiplex ELISA platform is highly effective in discrimination of infections with Bbss, B. garinii or B. afzelii. In the longer term, diagnostic peptides could be incorporated into a point of care rapid test.
Original language | English |
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Title of host publication | 16th International Conference on Lyme Borreliosis and other Tick-borne diseases; 4 to 7 September 2022 Amsterdam the Netherlands |
Publication status | Published - 5 Sept 2022 |
Externally published | Yes |
Event | 16th International Conference on Lyme Borreliosis and other Tick-borne diseases - Amsterdam, Netherlands Duration: 4 Sept 2022 → 7 Sept 2022 https://congresscare.com/en/congress/16th-international-conference-on-lyme-borreliosis-and-other-tick-borne-diseases-2/#:~:text=Tick%2Dborne%20diseases-,16th%20International%20Conference%20on%20Lyme%20Borreliosis%20and%20other%20Tick%2Dborne,place%20in%20Amsterdam%2C%20the%20Netherlands. |
Conference
Conference | 16th International Conference on Lyme Borreliosis and other Tick-borne diseases |
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Country/Territory | Netherlands |
City | Amsterdam |
Period | 4/09/22 → 7/09/22 |
Internet address |