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Metody používané pro identifikaci specifické DNA v klinickém materiálu

jsou shodné s následnými citacemi a komerčními kity.


 1.Borrelia bugdorferi sensu lato, Borrelia garinii, Borrelia afzelii, Borrelia burgdorferi sensu stricto – průkaz genu 16SrRNA genomové DNA.


ObrazekPrimery BG,VS,BB,LD, z genu 16S rRNA podukt  574,591,574, 356bp  teplota annealing 47 C




a)Richard T.Marconi and Claudie F.Garon

 Development of Polymerase Chain Reaction Primer Sets for Diagnosis of Lyme Disease and for Species-Specific Identification of Lyme Disease Isolates by 16S rRNA Signature Analysis.

Journal of Clinical Microbiology,Nov.1992,p.2830-2834


ObrazekPrimer 16S ribosom, produkt 169 bp, teplota annealing    55 C




b)Benjamin J.Luft,MD, Charles R.Steiman, MD, Harold C.Neimark,PhD., Bethi Muralidhar,PhD, Thomas Rush, MD, Michael F.Finkel, MD, Mark Kunkel, MD, Raymond J.Dattwyler,MD.


Invasion of the Central Nervous Systém by Borrelia burgdorferi in Acute Disseminated Infection

JAMA,March 11, 1992- Vol.267,No 10, p.1364-1367

 2.Borrelia burgdorferi sensu lato gen plasmidové DNA OSPA

 OspA-L5 a OspA-R5, produkt 445 bp, teplota annealing  68 C

 a)Mark M.Manak,PhD. Lucia I.González-Villaseňor,PhD.,Sylvia Crush-Stanton, and Richard C.Tilton,PhD

Use of PCR Assays to Monitor the Clearance of Borrelia burgdorferi DNA From Blood Following Antibiotic Therapy

Journal of Spirochetal and Tick-borne Diseases, Vol.4, No ½ Spring/Summer 1997 p.11-20






primer OspA 149  a OspA 319(P), produkt  194bp   teplota annealing  55 C


US Patent 6087097


b)Andrew R.Pachner, WeiFen Zhang, Henry Schaeffer, Susan Schaeffer and Tim O´Neill

Detection of Active Infection in Nonhuman Primates with Lyme Neuroborreliosis: Comparasion of PCR,Culture, and a Bioassay.

Journal of Clinical Microbiology, Nov. 1998, p.3243-3247


 Primery P66 1.krok  produkt 371 bp(inter.kontrola primer PDH 185 bp)tepl. annealing  42C

 Primery  P66 2.krok produkt 236 bp, tepl. annealing  .42 C

 c)Susanne Priem, Michael G.Rittig, Thomas Kamradt, Gerd E.Burmester and Andrea Krause,(An Optimized PCR Leads to Rapid and Highly Sensitive Detectionm od Borrelia burgdorferi in Patiens with Lyme Borreliosis.Journal of Clinical Mikrobiology, Mar.1997, p.685-690)

 d)Komerční kit pro DNA Borrelia burgdorferi

Genekam Biotechnology AG kat. K018

 3.Metody použité k identifikaci DNA Bartonella-specific

primer ITS 16S-23S  produkt 670 bp, teplota annealing  50 C

a)J M Rolain, F Gouriet, M Enea, M Aboud, D Raoult (2003)

Detection by immunofluorescence assay of Bartonella henselae in lymph nodes from patients with cat scratch disease.  

Clin Diagn Lab Immunol 10: 4. 686-691 Jul  

  primer ITS 16S-23S  produkt 670 bp, teplota 50 Cannealing 

b)Jennifer B. Henn,1,5 Mourad W. Gabriel,2,4 Rickie W. Kasten,1 Richard N. Brown,2 Jerold H. Theis,3 Janet E. Foley,4 and Bruno B. Chomel1

Gray Foxes (Urocyon cinereoargenteus) as a Potential Reservoir of a Bartonella clarridgeiae-Like Bacterium and Domestic Dogs as Part of a Sentinel System for Surveillance of Zoonotic Arthropod-Borne Pathogens in Northern California

 J Clin Microbiol. 2007 August; 45(8): 2411–2418.

 Primer  Bh16SF a Bh16SR produkt 185bp, teplota annealing  54 C

c)Edward B.Breitschwerdt, Barbara C.Hegarty and Susan I.HancockSequential Evaluation of Dogs Naturally Infectetd with Ehrlichia canis,Ehrlichia chaffeensis,Ehrlichia equi, Ehrlichia ewingii or Bartonella visonii 

Journal of Clinical Microbiology,Sept.1998,p2645-2651 Vol.36,No.9     

 4.Metody používané k identifikaci DNA Anaplasma phagocytophilum HGE


ObrazekPrimery 593II a ERB2 genu groEL   produkt 619 bp, teplota. annealing  65 C


a)Eva Olsson Engvall, Bertil Pettersson, Mariane Persson,Karin Artursson  and Karl-Erik Johansson.


A 16S rRNA-Based PCR Assay for Detection and Identification of Granulocytic Ehrlichia Species in Dogs, Horses, and Cattle.

Journal of Clinical Microbiology,Sept.1996,p 2170-2174 Vol.34,No.9


Primer EphplgroEL (569)F a EphplgroEL (1142) R produkt 573bp tep.ann.53 C

 Primer EphplgroEL (1193)R a EplgroEL (1084)R produkt 525bp tep.ann.67 C


Equine and Canine Anaplasma phagocytophilum Strains Isolated on the Island of Sardinia (Italy) Are Phylogenetically Related to Pathogenic Strains from the United States.Alberto Alberti,  Rosanna Zobba, Bernardo Chessa, Maria Filippa Addis , Oliver Sparagano, Maria Luisa Pinna Parpaglia, Tiziana Cubeddu, Gianpaolo Pitori,and Marco Pittau

Appl.Environ. Microbiol.2005, October, 71(10): 6418-6422


 c)Komerční kit k průklazu DNA Anaplasma phagocytophilum

Genekam Biotechnology AG kat. K097


 5.Metody používané k identifikaci DNA prvoka rodu Babesia.

 primer FOR 1 a REV 1 fragmentu genu 18S rRNA, produkt 1700 bp, teplota annealing  .54 C

 a)B.Skotarczak, M.Adamska, M.Supron

Blood DNA Analysis for Ehrlichia /Anaplasma)phagocytophila and Babesia spp. Of Dog from Northen Poland 

ACTA VET.BRNO 2004, 73,347-351

 b)komeční kit  pro DNA Babesia gibsoni

firma Genekam Biotechnology AG kat. K022


 c) komerční kit pro DNA Babesia bovis

firma Genekam Biotechnology AG kat. K078


6.Metody používané k indentifikaci DNA Rickettsií všech druhů mimo R.helvetica




Primery 120-M59 a 120-807 rOmpB gen, produkt 885bp. annealing  .teplota 50 C

a)V.Roux and D.Raoult

 Phylogenetic analysis of members of the genus Rickettsia using the gene encoding the outer-membrane protein rOmpB(ompB)

International Journal of Systematic and Evolutionary Microbiology (2000) 50,1449-1455

 b)Komerční kit pro DNA  Ricktettsia rickettsii

Genekam Biotechnology AG, kat. K713