Evaluation of Modified Two-Tiered Testing Algorithms for Lyme Disease Laboratory Diagnosis Using Well-Characterized Serum Samples

Adoracion Pegalajar-Jurado; Martin E Schriefer; Ryan J Welch; Marc R Couturier; Tiffany MacKenzie; Rebecca J Clark; Laura V Ashton; Mark J Delorey; Claudia R Molins

doi:10.1128/JCM.01943-17

Evaluation of Modified Two-Tiered Testing Algorithms for Lyme Disease Laboratory Diagnosis Using Well-Characterized Serum Samples

J Clin Microbiol. 2018 Jul 26;56(8):e01943-17. doi: 10.1128/JCM.01943-17. Print 2018 Aug.

Authors

Adoracion Pegalajar-Jurado¹, Martin E Schriefer¹, Ryan J Welch², Marc R Couturier^{2

3}, Tiffany MacKenzie⁴, Rebecca J Clark¹, Laura V Ashton¹, Mark J Delorey¹, Claudia R Molins⁵

Affiliations

¹ Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.
² ARUP Laboratories, Salt Lake City, Utah, USA.
³ Department of Pathology, University of Utah, Salt Lake City, Utah, USA.
⁴ DiaSorin, Inc., Stillwater, Minnesota, USA.
⁵ Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA ard5@cdc.gov.

Abstract

Standard two-tiered testing (STTT) is the recommended algorithm for laboratory diagnosis of Lyme disease (LD). Several limitations are associated with STTT that include low sensitivity in the early stages of disease, as well as technical complexity and subjectivity associated with second-tier immunoblotting; therefore, modified two-tiered testing (MTTT) algorithms that utilize two sequential first-tier tests and eliminate immunoblotting have been evaluated. Recently, a novel MTTT that uses a VlsE chemiluminescence immunoassay followed by a C6 enzyme immunoassay has been proposed. The purpose of this study was to evaluate the performance of the VlsE/C6 MTTT using well-characterized serum samples. Serum samples from the CDC Lyme Serum Repository were tested using three MTTTs, VlsE/C6, whole-cell sonicate (WCS)/C6, and WCS/VlsE, and three STTTs (immunoblotting preceded by three different first-tier assays: VlsE, C6, and WCS). Significant differences were not observed between the results of the MTTTs assessed; however, the VlsE/C6 MTTT resulted in the highest specificity (100%) when other diseases were tested and the lowest sensitivity (75%) for LD samples. Significant differences were present between the results for various MTTTs and STTTs evaluated. Specifically, all MTTTs resulted in higher sensitivities than the STTTs for all LD groups combined and were significantly more accurate (i.e., higher proportion of correct classifications) for this group, with the exception of the WCS/ViraStripe STTT. Additionally, when other diseases were tested, only the results of the VlsE/C6 MTTT differed significantly from those of the WCS/ViraStripe STTT, with the VlsE/C6 MTTT resulting in a 6.2% higher accuracy. Overall, the VlsE/C6 MTTT offers an additional laboratory testing algorithm for LD with equivalent or enhanced performance compared to that of the other MTTTs and STTTs evaluated in this study.

Keywords: Borrelia burgdorferi; Lyme disease; immunoserology; two-tiered testing.

Publication types

Comparative Study
Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Algorithms*
Antibodies, Bacterial / blood
Antigens, Bacterial / immunology
Bacterial Proteins / immunology
Borrelia burgdorferi / immunology*
Borrelia burgdorferi / isolation & purification
Humans
Immunoassay / standards*
Lipoproteins / immunology
Lyme Disease / blood
Lyme Disease / diagnosis*
Sensitivity and Specificity
Serologic Tests / standards*

Substances

Antibodies, Bacterial
Antigens, Bacterial
Bacterial Proteins
Lipoproteins
VlsE protein, Borrelia burgdorferi