Borrelia burgdorferi sensu lato (Borrelia burgdorferi sensu stricto, B. garinii, B. afzelii, B. lusitaniae, .... and other species) - Culture; IgG and IgG antibodies; specific antibodies (immunoblot); Molecular diagnosis (PCR); Molecular identification
Lyme borreliosis is a complex transmitted from animals (zoonosis), by Ixodes ticks, which act as vectors diseases. The most common zoonosis, transmitted by arthropods in warm areas of the Old and New World is considered. This zoonosis was named for Lyme disease or Lyme borreliosis, having been described in boxes arthritis in the 1970s, in the coastal district of "Old Lyme" in Connecticut (USA). Causing tick - borne spirochetes were included in the genus Borrelia, and named Borrelia burgdorferi. Subsequently they were showing differences among isolates of Borrelia for various types of related processes, and were differentiated into species. Therefore now considered a group of species collectively known as Borrelia burgdorferi sensu lato (broadly defined). Furthermore, it is known that reservoirs vary from species to species, and that the geographical distribution also varies. The reservoir also influences and explains the genetic diversity that can be found for the same species. When they are migratory birds, genetic heterogeneity is superior to that possessed species whose reservoir is in mammals, reptiles or which are more homogeneous.
In Lyme Borreliosis, the initial sign is a pox lesion Erythema migrans call, on the site of the bite of the tick transmitting. After inoculation infection may affect other organs, which is considered producing a multisystem disease even though there is not existed the initial injury Erythema migrans, affecting joints (Lyme arthritis), central nervous system (neuroborreliosis ), and chronic cutaneous manifestations (Acrodermatitis chronic atrophic). For infection to manifest great diversity of clinical manifestations, it has been called the "great imitator".
The joint (Lyme arthritis), manifestation is characterized by muscle and joint symptoms, and is preferably produced by Borrelia burgdorferi sensu stricto. In U.S.A. 60% of infected patients who are not treated develop joint demonstration. However, in Europe the articular manifestation is less frequent (3 to 15% of infections), because the species Borrelia afzelii found are preferably garinii and Borrelia.
Neuroborreliosis, is preferably produced by Borrelia garinii, but can also be produced by Borrelia garinii and Borrelia burgdorferi sensu stricto.
Skin manifestations (Erythema migrans, acrodermatitis chronica atrophicans and benign skin Lymphocytoma) are preferably produced by Borrelia afzelii, but can also be produced by other species.
Species (genospecies) currently supported within the complex Borrelia burgdorferi sensu lato, related clinical conditions are as follows (9 species):
• Borrelia burgdorferi sensu stricto, is the most important species in the US It is in Eurasia, in Europe, including the Iberian Peninsula, but not in Asia. It has clear pathogenic significance. It is the only one of the three classic pathogenic species (Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii), located in the US and Eurasia.
• Borrelia garinii, reservoir in birds, is in Eurasia, including the Iberian Peninsula. It has clear pathogenic significance.
• Borrelia afzelii, reservoir in rodents, is in Eurasia, including the Iberian Peninsula. It has clear pathogenic significance.
• Borrelia lusitaniae, reservoir lizards (Lacertidae family), is transmitted by Ixodes ricinus tick species. It is located in Eurasia, as in the central and northern Portugal and Spain, along with other species, and as far as is known in southern Portugal and North Africa as a unique species. It described as causing a chronic skin lesion in a 46 years in Portugal.
• Borrelia valaisiana, reservoir in birds. It is located in Eurasia, and the Iberian Peninsula.
• Borrelia bavariensis. It is located in Eurasia.
• Borrelia bisettii. It is located in the US and Eurasia.
• Borrelia kurtenbachii. It is located in the US
• Borrelia spielmanii. It is located in Eurasia.
No pathogenic species, or at least not yet detected, are as follows (9 species):
• American Borrelia. It is located in the US
• Borrelia andersonii. It is located in the US
• Borrelia californiensis. It is located in the US
• Borrelia carolinensis. It is located in the US and Eurasia.
• Borrelia japonica. It is located in Eurasia.
• Borrelia tanukii. It is located in Eurasia.
• Borrelia Turdi. It is located in Eurasia.
• Borrelia sinica. It is located in Eurasia.
• Borrelia yangtse. It is located in Eurasia.
Microbiological diagnostic methods:
• Isolation in culture: the most recommended is the medium of Barbour-Stoenuer-Kelly, but has the disadvantage that a long incubation period of up to 4 weeks is required with a weekly microscopic examination to see whether there has been growing .
• Detection and molecular differentiation of species by PCR, with or without sequencing: This method offers the advantage of speed, and if the common and differential molecular target between species of Borrelia burgdorferi sensu lato is used. It differentiates to what species we are, by sequencing.
• Detection of total antibodies, IgG antibodies or IgM antibodies: this test is useful as usually the diagnosis of Lyme borreliosis is a late diagnosis, and have been induced antibodies. The problem is that sometimes IgG antibodies or IgM class detected by an immunosorbent assay (ELISA) or immunofluorescence may give false positive results to be cross - react with other spirochetes (Treponema pallidum, Leptospira spp.).
• Detection of antibodies to specific antigens by Western Blot: to differentiate cross - reactive antibodies, specific antibodies generated against Borrelia burgdorferi sensu lato, or against some species, is required to perform a procedure type Immunoblot -Western- Line blot or assay-, using highly purified antigens Borrelia burgdorferi sensu lato (OspA, OspC, p100, VlsE, p39, p58 [DbpA, decorin-binding protein a], p41 [flagellin]), separated by electrophoresis and transferred to a nitrocellulose membrane; or use recombinant antigens in "Line assay" type method. Thus they can detect specific antibodies against one / s / of the antigen / s of Borrelia burgdorferi sensu lato specific or some specific species of Borrelia, Borrelia garinii and Borrelia afzelii, Borrelia bavariensis, and Borrelia spielmanii.
Tests in IVAMI:
• Isolation in culture.
• Molecular detection by PCR.
• Differentiation causative species by PCR, followed by sequencing the amplicon obtained.
• Detection of total antibodies, IgM or IgG antibodies by enzyme immunoassay (ELISA) for IgM and / or IgG.
• Detection of antibodies to specific antigens Borrelia by immunoblot, both serum and cerebrospinal fluid.
Sample / s recommended / s:
Detection of current infection with Borrelia burgdorferi sensu lato
• Skin Biopsies in cases Eryhthema migrans or Dermatitis Chronic Atrophic biopsy deposited in a sterile tube sealing, preferably polypropylene to avoid breakage during transport to the laboratory.
• central nervous system in cases of neuroborreliosis: cerebrospinal fluid sample, 1 to 2 mL in sterile polypropylene vial.
Detection of current or past infection by antibodies
• Whey: sample 1 to 2 mL serum vial in sterile polypropylene.
• Cerebrospinal fluid: sample 1 to 2 mL serum vial in sterile polypropylene.
Preservation and shipment of sample:
Refrigerated (preferred) for less than 2 days.
Frozen: over 2 days.
Delivery times :
• Isolation in culture: up to 4 weeks.
• Molecular detection by PCR: 48 to 72 hours
• Differentiation causative species by PCR, followed by sequencing the amplicon obtained 48 to 72 hours.
• Detection of total antibodies, IgM or IgG antibodies by enzyme immunoassay (ELISA) for IgM and / or IgG: 48 to 72 hours.
• Detection of antibodies to specific antigens Borrelia by immunoblot: 48 to 72 hours.
Cost of testing: