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Detection of Borreliacidal Antibodies by Flow Cytometry: An Accurate, Highly Specific Serodiagnostic Test for Lyme Disease

Detection of Borreliacidal Antibodies by Flow Cytometry: An Accurate, Highly Specific... Abstract Background: Borreliacidal antibodies can be detected in serum samples from patients with early or late Lyme disease symptoms. When these serum samples are incubated with Borrelia burgdorferi and complement, spirochetes are rapidly killed. Detection of these antibodies can be used as a serodiagnostic test. Methods: Individual serum samples containing IgM or IgG borreliacidal antibodies were used to develop a method for detection using flow cytomety. An additional 10 case-defined Lyme disease serum samples and 10 normal serum samples were used to confirm appropriate flow cytometric parameters. To determine specificity, 157 normal serum samples and 104 potential cross-reactive serum samples were tested for borreliacidal activity and antibodies to B burgdorferi using indirect fluorescent antibody or enzyme immunoassay. Results: Flow cytometry can be used to detect borreliacidal activity within 16 to 24 hours after incubation of B burgdorferi organisms, Lyme disease serum, and complement. Significant borreliacidal activity was detected in all Lyme disease serum samples. The percentages of positive normal serum samples were comparable (6% to 10%) using all three assays. In addition, the indirect fluorescent antibody and enzyme immunoassay identified 41 (39%) and 47 (45%) potential cross-reactive serum samples as positive, respectively. In contrast, significant borreliacidal activity was not detected in any potential cross-reactive serum samples. Conclusion: Detection of borreliacidal antibody, unlike indirect fluorescent antibody and enzyme immunoassay, is an accurate, highly specific serodiagnostic test for detection of Lyme disease.(Arch Intern Med. 1994;154:1625-1632) References 1. Steere AC, Grodzicki RL, Kornblatt AN, et al. The spirochetal etiology of Lyme disease . N Engl J Med . 1983;308:733-740.Crossref 2. Centers for Disease Control and Prevention. Lyme disease surveillance— United States, 1989-1990 . MMWR Morb Mortal Wkly Rep . 1991;40:417-420. 3. Steere AC, Bartenhagen NH, Craft JE, et al. The early clinical manifestations of Lyme disease . Ann Intern Med . 1983;99:76-82.Crossref 4. Pachner AR, Steere AC. The triad of neurologic manifestations of Lyme disease: meningitis, cranial neuritis, and radiculoneuritis . Neurology . 1985;35:47-53.Crossref 5. Steere AC, Batsford WP, Weinberg M, et al. Lyme carditis: cardiac abnormalities of Lyme disease . Ann Intern Med . 1980;93:8-16.Crossref 6. Steere AC, Schoen RT, Taylor E. The clinical evolution of Lyme arthritis . Ann Intern Med . 1987;107:725-731.Crossref 7. Agger WA, Case KL, Bryant GL, Callister SM. Lyme disease: clinical features, classification, and epidemiology in the upper midwest . Medicine . 1991;70:83-90.Crossref 8. Hedberg CW, Osterholm MT, MacDonald KL, White KE. An interlaboratory study of antibody to Borrelia burgdorferi . J Infect Dis . 1987;155:1325-1327.Crossref 9. Bakken LL, Case KL, Callister SM, Bourdeau NJ, Schell RF. Performance of 45 laboratories participating in a proficiency testing program for Lyme disease serology . JAMA . 1992;268:891-895.Crossref 10. Steere AC, Taylor E, McHugh GL, Logigian EL. The overdiagnosis of Lyme disease . JAMA . 1993;269:1812-1816.Crossref 11. Hedberg CW, Osterholm MT. Serologic tests for antibody to Borrelia burgdorferi: another Pandora's box for medicine? Arch Intern Med . 1990;150:732-733.Crossref 12. Lane RS, Lennette ET, Madigan JE. Interlaboratory and intralaboratory comparisons of indirect immunofluorescence assays for serodiagnosing Lyme disease . J Clin Microbiol . 1990;28:1774-1779. 13. Luger SW, Krauss E. Serologic tests for Lyme disease: interlaboratory variability . Arch Intern Med . 1990;150:761-763.Crossref 14. Schwartz BS, Goldstein MD, Ribiero JMC, Schulze TL, Shaheid SI. Antibody testing in Lyme disease: a comparison of results in four laboratories . JAMA . 1989;262:3431-3434.Crossref 15. Jones JM. Serodiagnosis of Lyme disease . Ann Intern Med . 1991;114:1064.Crossref 16. Bakken L, Case K, Schell R, Homburger H, Bourdeau N, Callister SM. Variability of 500 laboratories participating in a proficiency program (WSLH/CAP) for Lyme disease serodiagnosis . Abstracted from the American Society for Microbiology General Meeting, Atlanta, Ga, 1993 . 17. Callister SM, Schell RF, Lovrich SD. Lyme disease assay which detects killed Borrelia burgdorferi . J Clin Microbiol . 1991;29:1773-1776. 18. Callister SM, Schell RF, Case KL, Lovrich SD, Day SP. Characterization of the borreliacidal antibody response to Borrelia burgdorferi in humans: a serodiagnostic test . J Infect Dis . 1993;167:158-164.Crossref 19. Barbour AG. Isolation and cultivation of Lyme disease spirochetes . Yale J Biol Med . 1984;57:521-525. 20. Callister SM, Case KL, Agger WA, Schell RF, Johnson RC, Ellingson JLE. Effects of bovine serum albumin on the ability of Barbour-Stoenner-Kelly medium to detect Borrelia burgdorferi . J Clin Microbiol . 1990;28:363-365. 21. Sachsenmeier KF, Schell K, Morrissey LW, et al. Detection of borreliacidal antibodies in hamsters by using flow cytometry . J Clin Microbiol . 1992;30:1457-1461. 22. Lim LCL, Liu YF, Schell K, Lovrich SD, Callister SM, Schell RF. Detection of borreliacidal antibody by using acridine orange and flow cytometry . Clin Diagn Lab Immunol . 1994;1:44-50. 23. Chapin-Robertson K, Edberg SC. Measurement of antibiotics in human body fluids: techniques and significance . In: Lorian V, ed. Antibiotics in Laboratory Medicine . 3rd ed. Baltimore, Md: Williams & Wilkins; 1991:295-366. 24. Russell H, Sampson JS, Schmid GP, Wilkinson HW, Plikaytis B. Enzyme-linked immunosorbent assay and indirect immunofluorescence assay for Lyme disease . J Infect Dis . 1984;149:465-470.Crossref 25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent . J Biol Chem . 1951;193:265-275. 26. Craft JE, Grodzicki RL, Steere AC. Antibody response in Lyme disease: evaluation of diagnostic tests . J Infect Dis . 1984;149:789-795.Crossref 27. Steel RGD, Torrie JH. Principles and Procedures of Statistics With Special References to the Biological Sciences . New York, NY: McGraw-Hill International Book Co; 1960:67-87. 28. Lovrich SD, Callister SM, Schmitz JL, Alder JF, Schell RF. Borreliacidal activity of sera from hamsters infected with the Lyme disease spirochete . Infect Immun . 1991;59:2522-2528. 29. Schmitz JL, Schell RF, Heyka AG, England DM. Passive immunization prevents induction of Lyme arthritis in LSH hamsters . Infect Immun . 1990;58: 144-148. 30. Pavia CS, Kissel V, Bitker S, Cabello F, Levine S. Anti-borrelial activity of rats injected with the Lyme disease spirochete . J Infect Dis . 1991;163:165-169.Crossref 31. Philipp MT, Aydintug MK, Bohm RP Jr, et al. Early and early disseminated phases of Lyme disease in the rhesus monkey: a model for infection in humans . Infect Immun . 1993;61:3047-3059. 32. Schaible EU, Kramer MD, Eichmann K, Modolell M, Museteanu C, Simon MM. Monoclonal antibodies specific for the outer protein (OspA) of Borrelia burgdorferi prevent Lyme borreliosis in severe combined immunodeficiency mice . Proc Natl Acad Sci U S A . 1990;87:3768-3772.Crossref 33. Fikrig E, Barthold SW, Kantor FS, Flavell RA. Protection of mice against the Lyme disease agent by immunizing with recombinant OspA . Science . 1990; 250:553-555.Crossref 34. Fikrig E, Barthold SW, Kantor FS, Flavell RA. Long-term protection of mice from Lyme disease by vaccination with OspA . Infect Immun . 1992;60:773-777. 35. Johnson RC, Kodner C, Russell M. Passive immunization of hamsters against experimental infection with the Lyme disease spirochete . Infect Immun . 1986; 53:713-714. 36. Chu HJ, Chavez LG, Blumer BM, Sebring RW, Wasmoen TL, Acree WM. Immunogenicity and efficacy study of a commercial Borrelia burgdorferi bacteria . J Am Vet Med Assoc . 1992;201:403-411. 37. Sadziene A, Thompson PA, Barbour AG. In vitro inhibition of Borrelia burgdorferi growth by antibodies . J Infect Dis . 1993;167:165-172.Crossref 38. Heyka A, Schmitz JL, England DE, Callister SM, Schell RF. Histopathology of Lyme arthritis in LSH hamsters . Am J Pathol . 1989;134:1113-1123. 39. Duray PH, Johnson RC. The histopathology of experimentally infected hamsters with the Lyme disease spirochete, Borrelia burgdorferi (42251) . Proc Soc Exp Biol Med . 1986;181:263-269.Crossref 40. Johnson RC, Kodner C, Russell M, Duray PH. Experimental infection of the hamster with Borrelia burgdorferi . Ann N Y Acad Sci . 1988;539:258-263.Crossref 41. Barbour AG, Hayes SF. Biology of Borrelia species . Microbiol Rev . 1986;50: 381-400. 42. Coleman JL, Benach JL. Characterization of antigenic determinants of Borrelia burgdorferi shared by other bacteria . J Infect Dis . 1992;165:658-666.Crossref 43. Magnarelli LA, Miller JM, Anderson JF, Riviere GF. Cross-reactivity of nonspecific treponemal antibody in serologic tests for Lyme disease . J Clin Microbiol . 1990;28:1276-1279. 44. Garcia-Monco JC, Coleman JL, Benach JL. Antibodies to myelin basic protein in Lyme disease . J Infect Dis . 1988;158:667-668.Crossref 45. Duffy J, Mertz LE, Wobig GH, Katzmann JA. Diagnosing Lyme disease: the contribution of serologic testing . Mayo Clin Proc . 1988;63:1116-1121.Crossref 46. Lovrich SD, Callister SM, Lim LCL, Schell RF. Seroprotective groups among isolates of Borrelia burgdorferi . Infect Immun . 1993;61:4367-4374. 47. Fikrig E, Barthold SW, Marcantoni N, Deponti K, Kantor FS, Flavell RA. Role of Osp A, Osp B, and flagellin in protective immunity of Lyme borreliosis in the laboratory mouse . Infect Immun . 1992;60:657-661. 48. Preac-Mursic V, Wilske B, Patsouris E, et al. Active immunization with pC protein of Borrelia burgdorferi protects gerbils against Borrelia burgdorferi infection . Infection . 1992;20:342-349.Crossref 49. Scriba M, Ebrahim JL, Schlott T, Eiffert H. The 39 kilodalton protein of Borrelia burgdorferi: a target for bactericidal human monoclonal antibodies . Infect Immun . 1993;61:4523-4526. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Internal Medicine American Medical Association

Detection of Borreliacidal Antibodies by Flow Cytometry: An Accurate, Highly Specific Serodiagnostic Test for Lyme Disease

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Publisher
American Medical Association
Copyright
Copyright © 1994 American Medical Association. All Rights Reserved.
ISSN
0003-9926
eISSN
1538-3679
DOI
10.1001/archinte.1994.00420140095012
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Abstract

Abstract Background: Borreliacidal antibodies can be detected in serum samples from patients with early or late Lyme disease symptoms. When these serum samples are incubated with Borrelia burgdorferi and complement, spirochetes are rapidly killed. Detection of these antibodies can be used as a serodiagnostic test. Methods: Individual serum samples containing IgM or IgG borreliacidal antibodies were used to develop a method for detection using flow cytomety. An additional 10 case-defined Lyme disease serum samples and 10 normal serum samples were used to confirm appropriate flow cytometric parameters. To determine specificity, 157 normal serum samples and 104 potential cross-reactive serum samples were tested for borreliacidal activity and antibodies to B burgdorferi using indirect fluorescent antibody or enzyme immunoassay. Results: Flow cytometry can be used to detect borreliacidal activity within 16 to 24 hours after incubation of B burgdorferi organisms, Lyme disease serum, and complement. Significant borreliacidal activity was detected in all Lyme disease serum samples. The percentages of positive normal serum samples were comparable (6% to 10%) using all three assays. In addition, the indirect fluorescent antibody and enzyme immunoassay identified 41 (39%) and 47 (45%) potential cross-reactive serum samples as positive, respectively. In contrast, significant borreliacidal activity was not detected in any potential cross-reactive serum samples. Conclusion: Detection of borreliacidal antibody, unlike indirect fluorescent antibody and enzyme immunoassay, is an accurate, highly specific serodiagnostic test for detection of Lyme disease.(Arch Intern Med. 1994;154:1625-1632) References 1. Steere AC, Grodzicki RL, Kornblatt AN, et al. The spirochetal etiology of Lyme disease . N Engl J Med . 1983;308:733-740.Crossref 2. Centers for Disease Control and Prevention. Lyme disease surveillance— United States, 1989-1990 . MMWR Morb Mortal Wkly Rep . 1991;40:417-420. 3. Steere AC, Bartenhagen NH, Craft JE, et al. The early clinical manifestations of Lyme disease . Ann Intern Med . 1983;99:76-82.Crossref 4. Pachner AR, Steere AC. The triad of neurologic manifestations of Lyme disease: meningitis, cranial neuritis, and radiculoneuritis . Neurology . 1985;35:47-53.Crossref 5. Steere AC, Batsford WP, Weinberg M, et al. Lyme carditis: cardiac abnormalities of Lyme disease . Ann Intern Med . 1980;93:8-16.Crossref 6. Steere AC, Schoen RT, Taylor E. The clinical evolution of Lyme arthritis . Ann Intern Med . 1987;107:725-731.Crossref 7. Agger WA, Case KL, Bryant GL, Callister SM. Lyme disease: clinical features, classification, and epidemiology in the upper midwest . Medicine . 1991;70:83-90.Crossref 8. Hedberg CW, Osterholm MT, MacDonald KL, White KE. An interlaboratory study of antibody to Borrelia burgdorferi . J Infect Dis . 1987;155:1325-1327.Crossref 9. Bakken LL, Case KL, Callister SM, Bourdeau NJ, Schell RF. Performance of 45 laboratories participating in a proficiency testing program for Lyme disease serology . JAMA . 1992;268:891-895.Crossref 10. Steere AC, Taylor E, McHugh GL, Logigian EL. The overdiagnosis of Lyme disease . JAMA . 1993;269:1812-1816.Crossref 11. Hedberg CW, Osterholm MT. Serologic tests for antibody to Borrelia burgdorferi: another Pandora's box for medicine? Arch Intern Med . 1990;150:732-733.Crossref 12. Lane RS, Lennette ET, Madigan JE. Interlaboratory and intralaboratory comparisons of indirect immunofluorescence assays for serodiagnosing Lyme disease . J Clin Microbiol . 1990;28:1774-1779. 13. Luger SW, Krauss E. Serologic tests for Lyme disease: interlaboratory variability . Arch Intern Med . 1990;150:761-763.Crossref 14. Schwartz BS, Goldstein MD, Ribiero JMC, Schulze TL, Shaheid SI. Antibody testing in Lyme disease: a comparison of results in four laboratories . JAMA . 1989;262:3431-3434.Crossref 15. Jones JM. Serodiagnosis of Lyme disease . Ann Intern Med . 1991;114:1064.Crossref 16. Bakken L, Case K, Schell R, Homburger H, Bourdeau N, Callister SM. Variability of 500 laboratories participating in a proficiency program (WSLH/CAP) for Lyme disease serodiagnosis . Abstracted from the American Society for Microbiology General Meeting, Atlanta, Ga, 1993 . 17. Callister SM, Schell RF, Lovrich SD. Lyme disease assay which detects killed Borrelia burgdorferi . J Clin Microbiol . 1991;29:1773-1776. 18. Callister SM, Schell RF, Case KL, Lovrich SD, Day SP. Characterization of the borreliacidal antibody response to Borrelia burgdorferi in humans: a serodiagnostic test . J Infect Dis . 1993;167:158-164.Crossref 19. Barbour AG. Isolation and cultivation of Lyme disease spirochetes . Yale J Biol Med . 1984;57:521-525. 20. Callister SM, Case KL, Agger WA, Schell RF, Johnson RC, Ellingson JLE. Effects of bovine serum albumin on the ability of Barbour-Stoenner-Kelly medium to detect Borrelia burgdorferi . J Clin Microbiol . 1990;28:363-365. 21. Sachsenmeier KF, Schell K, Morrissey LW, et al. Detection of borreliacidal antibodies in hamsters by using flow cytometry . J Clin Microbiol . 1992;30:1457-1461. 22. Lim LCL, Liu YF, Schell K, Lovrich SD, Callister SM, Schell RF. Detection of borreliacidal antibody by using acridine orange and flow cytometry . Clin Diagn Lab Immunol . 1994;1:44-50. 23. Chapin-Robertson K, Edberg SC. Measurement of antibiotics in human body fluids: techniques and significance . In: Lorian V, ed. Antibiotics in Laboratory Medicine . 3rd ed. Baltimore, Md: Williams & Wilkins; 1991:295-366. 24. Russell H, Sampson JS, Schmid GP, Wilkinson HW, Plikaytis B. Enzyme-linked immunosorbent assay and indirect immunofluorescence assay for Lyme disease . J Infect Dis . 1984;149:465-470.Crossref 25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent . J Biol Chem . 1951;193:265-275. 26. Craft JE, Grodzicki RL, Steere AC. Antibody response in Lyme disease: evaluation of diagnostic tests . J Infect Dis . 1984;149:789-795.Crossref 27. Steel RGD, Torrie JH. Principles and Procedures of Statistics With Special References to the Biological Sciences . New York, NY: McGraw-Hill International Book Co; 1960:67-87. 28. Lovrich SD, Callister SM, Schmitz JL, Alder JF, Schell RF. Borreliacidal activity of sera from hamsters infected with the Lyme disease spirochete . Infect Immun . 1991;59:2522-2528. 29. Schmitz JL, Schell RF, Heyka AG, England DM. Passive immunization prevents induction of Lyme arthritis in LSH hamsters . Infect Immun . 1990;58: 144-148. 30. Pavia CS, Kissel V, Bitker S, Cabello F, Levine S. Anti-borrelial activity of rats injected with the Lyme disease spirochete . J Infect Dis . 1991;163:165-169.Crossref 31. Philipp MT, Aydintug MK, Bohm RP Jr, et al. Early and early disseminated phases of Lyme disease in the rhesus monkey: a model for infection in humans . Infect Immun . 1993;61:3047-3059. 32. Schaible EU, Kramer MD, Eichmann K, Modolell M, Museteanu C, Simon MM. Monoclonal antibodies specific for the outer protein (OspA) of Borrelia burgdorferi prevent Lyme borreliosis in severe combined immunodeficiency mice . Proc Natl Acad Sci U S A . 1990;87:3768-3772.Crossref 33. Fikrig E, Barthold SW, Kantor FS, Flavell RA. Protection of mice against the Lyme disease agent by immunizing with recombinant OspA . Science . 1990; 250:553-555.Crossref 34. Fikrig E, Barthold SW, Kantor FS, Flavell RA. Long-term protection of mice from Lyme disease by vaccination with OspA . Infect Immun . 1992;60:773-777. 35. Johnson RC, Kodner C, Russell M. Passive immunization of hamsters against experimental infection with the Lyme disease spirochete . Infect Immun . 1986; 53:713-714. 36. Chu HJ, Chavez LG, Blumer BM, Sebring RW, Wasmoen TL, Acree WM. Immunogenicity and efficacy study of a commercial Borrelia burgdorferi bacteria . J Am Vet Med Assoc . 1992;201:403-411. 37. Sadziene A, Thompson PA, Barbour AG. In vitro inhibition of Borrelia burgdorferi growth by antibodies . J Infect Dis . 1993;167:165-172.Crossref 38. Heyka A, Schmitz JL, England DE, Callister SM, Schell RF. Histopathology of Lyme arthritis in LSH hamsters . Am J Pathol . 1989;134:1113-1123. 39. Duray PH, Johnson RC. The histopathology of experimentally infected hamsters with the Lyme disease spirochete, Borrelia burgdorferi (42251) . Proc Soc Exp Biol Med . 1986;181:263-269.Crossref 40. Johnson RC, Kodner C, Russell M, Duray PH. Experimental infection of the hamster with Borrelia burgdorferi . Ann N Y Acad Sci . 1988;539:258-263.Crossref 41. Barbour AG, Hayes SF. Biology of Borrelia species . Microbiol Rev . 1986;50: 381-400. 42. Coleman JL, Benach JL. Characterization of antigenic determinants of Borrelia burgdorferi shared by other bacteria . J Infect Dis . 1992;165:658-666.Crossref 43. Magnarelli LA, Miller JM, Anderson JF, Riviere GF. Cross-reactivity of nonspecific treponemal antibody in serologic tests for Lyme disease . J Clin Microbiol . 1990;28:1276-1279. 44. Garcia-Monco JC, Coleman JL, Benach JL. Antibodies to myelin basic protein in Lyme disease . J Infect Dis . 1988;158:667-668.Crossref 45. Duffy J, Mertz LE, Wobig GH, Katzmann JA. Diagnosing Lyme disease: the contribution of serologic testing . Mayo Clin Proc . 1988;63:1116-1121.Crossref 46. Lovrich SD, Callister SM, Lim LCL, Schell RF. Seroprotective groups among isolates of Borrelia burgdorferi . Infect Immun . 1993;61:4367-4374. 47. Fikrig E, Barthold SW, Marcantoni N, Deponti K, Kantor FS, Flavell RA. Role of Osp A, Osp B, and flagellin in protective immunity of Lyme borreliosis in the laboratory mouse . Infect Immun . 1992;60:657-661. 48. Preac-Mursic V, Wilske B, Patsouris E, et al. Active immunization with pC protein of Borrelia burgdorferi protects gerbils against Borrelia burgdorferi infection . Infection . 1992;20:342-349.Crossref 49. Scriba M, Ebrahim JL, Schlott T, Eiffert H. The 39 kilodalton protein of Borrelia burgdorferi: a target for bactericidal human monoclonal antibodies . Infect Immun . 1993;61:4523-4526.

Journal

Archives of Internal MedicineAmerican Medical Association

Published: Jul 25, 1994

References