|
Erreger
|
PCR-Methode
(einfach oder "nested PCR") |
PCR-Primer |
PCR-Bedingungen
(Annealing
Temperatur / Zyklen) |
Literatur
|
|
Borrelia burgdorferi |
nested PCR |
1. PCR:
(PCR-Produkt: 259 bp)
Bor-1:
5´-AGAAGTGCTGGAGTCGAAGCGAAA-3´
Bor-2:
5´-TAGTGCTCTACCTCTATTAAACTA-3´
2. PCR:
(PCR-Produkt: 219 bp)
Bor-3:
5´-GCGAAAGCGAGTCTTAAAAGG-3´
Bor-4:
5´-ACTAAAATAAGGCTGAACTTAAAT-3´ |
53 Grad / 40
Zyklen
58 Grad / 40 Zyklen |
(1) |
|
Treponema pallidum |
nested PCR |
1. PCR:
(PCR-Produkt: 379 bp)
Trep-1:
5´-CAGCAGGGGAAGAAAAAAGTGGG-3´
Trep-2:
5´-AAGGTCGTGCGGGCTCTCCAT-3´
2. PCR:
(PCR-Produkt: 196 bp)
Trep-3:
5´-GACCCAAGCGTTACTAAGATGG-3´
Trep-4: 5´-
ACCGCAACTGGGACAAACTTCAT-3´ |
66 Grad / 35
Zyklen
66 Grad / 35 Zyklen |
(2) |
|
Mykobakterien (Genus-spezifisch) |
nested PCR |
PCR-1:
(PCR-Produkt:
234 bp)
65kD-1:
5´-AGGCGTTGGTTCGCGAGGG-3´
65kD-2: 5´-GATGACGCCCTCGTTGCC-3´
PCR-2:
(PCR-Produkt 143 bp)
65 kD-3:
5´-CCAACCCGCTCGGTCTAA-3´
64
kD-4: 5´-CCGATGGACTGGTCACCC-3´ |
65°C / 40 Zyklen
65°C / 40 Zyklen |
(3) |
|
Mycobacterium leprae |
nested PCR |
PCR-1:
(PCR-Produkt: 372 bp)
LEP-1:
5´-GCACGTAAGCCTGTCGGTGG-3´
LEP-2:
5´-CGGCCGGATCCTCGATGCAC-3´
PCR-2:
(PCR-Produkt: 329 bp)
LEP-3:
5´-TGGGTGCTGCTTGGTCTACA-3´
LEP-4:
5´-TCACTAACACGATACTGCTG-3´ |
70 Grad / 40
Zyklen
62 Grad / 40 Zyklen |
(4) |
Humane Papillom Viren
(allgemein) |
nested PCR |
PCR-1:
(PCR-Produkt: 449 bp)
HPV-1:
5´-CGTCC(AC)A(AG)(AG)GGA(AT)ACTGATC-3´
HPV-2:
5´-GC(AC)CAGGG(AT)CATAA(CT)AATGG-3´
PCR-2:
(PCR-Produkt: 139 bp)
GP-5:
5´-TTTGTTACTGTGGTAGATAC-3´
GP-6:
5´-GAAAAATAAACTGTAAATCA-3´ |
46 Grad / 45
Zyklen
52 Grad / 45 Zyklen |
(5) |
Humane Papillom Viren
(HPV-7) |
einfach |
(PCR-Produkt 113 bp)
HPV-7:5´-AAATTTAACATTATGTGCTG-3´
GP6: 5´-GAAAATAAACTGTAAATCA-3´ |
52 Grad / 45
Zyklen |
(14) |
Humane Papillom Viren
(HPV-16) |
einfach |
(PCR-Produkt
342 bp)
HPV-16-F:
5´-GCAAACCACCTATAGGGGAA-3´
HPV-16-R:
5´-AATCGTCTGGTACATTTTCA-3´ |
61 Grad / 45
Zyklen |
(13) |
Humane Papillom Viren
(HPV-18) |
einfach |
(PCR-Produkt: 217 bp)
HPV-18-F:
5´-AAGGATGCTGCACCGGCTGA-3´
HPV-18-R 5´-CACGCACACGCTTGGCAGGT-3´ |
65 Grad / 45
Zyklen |
|
|
Cytomegalie-Virus (CMV) |
nested PCR |
1.PCR (PCR-Produkt 214 bp)
CMV-1: 5´-GGCCATCGCCGAGGAGTCA-3´
CMV-2: 5´-CCGCTCCTCCTGAGCACCC-3´
2. PCR
(PCR-Produkt 120 bp)
CMV-3:
5´-TCTGATTCTCTGGTGTCAC-3´
CMV-4: 5´-TCCTCCTCTTCCTCATCACT-3´ |
58 Grad / 45 Zyklen
50 Grad / 45 Zyklen
|
(16)
|
|
Epstein Barr Virus |
einfach |
(PCR-Produkt:
172 bp) EBV-1:
5´-CGCAGGGATGCCTGGACACA-3´
EBV-2:
5´-CTGTTTGTGGGCCTTGTGGC-3´ |
68 Grad / 40
Zyklen |
(6) |
Herpes Simplex Virus
Typ I+II |
einfach |
(HSV I-PCR-Produkt: 395 bp, HSV
II-PCR-Produkt: 302 bp)
HSV-F:
5´-CCGTGGCCCTGTGGGGCCTG-3´
HSV-R:
5´-CACCCGCGAGCCGTACCGGG-3´ |
74 Grad / 40
Zyklen |
(7) |
|
Humanes Herpesvirus Typ7 (HHV-7) |
einfach |
(PCR-Produkt: 186 bp)
HHV-7-1: 5´-TATCCCAGCTGTTTTCATATAGTAAC-3´
HHV-7-2: 5´-GCCTTGCGGTAGCACTAGATTTTTTG-3´ |
54 Grad / 45
Zyklen |
|
Kaposi Sarkom asso-ziierter
Herpes Virus
(KSHV / HHV-8) |
einfach |
(PCR-Produkt
234 bp) KSHV-1:
5´-AGCCGAAAGGATTCCACCAT-3´
KSHV-2:
5´-TCCGTGTTGTCTACGTCCAG-3´ |
58 Grad / 40
Zyklen |
(8)
|
|
Varicella Zoster Virus |
einfach |
(PCR-Produkt
224 bp) VZV-1:
5´-CGTCACATATTATGCAAACATG-3´
VZV-2:
5´-CGTTTTTAATATTACAAATCCCGC-3´ |
58 Grad / 40
Zyklen |
(9)
|
|
Parvovirus B 19 |
einfach |
(PCR-Produkt
422 bp) B19-1:
5´-TTCTTTTCAGCTTTTAGG-3´
B19-2:
5´-GTACTTCTGGTACGTTAAGT-3´ |
Touchdown von 65 Grad nach 55 Grad, bei 55 Grad 35 Zyklen |
(10) |
|
Leishmanien |
nested PCR |
PCR-1:
LEI-1:
5´-CCAACTGGGGGTTGGTGTAA-3´
LEI-2:
5´-TTTTGAACGGGGTTTCTG-3´
PCR-2:
(PCR-Produkt
je nach Art 280-415 bp)
LEI-3: GGTATGCGAAACTTCCGGAGA
LEI-4: GTATACTTATATAGCGTTAG |
48 Grad / 45
Zyklen
52 Grad / 45 Zyklen |
(11) |
|
Rickettsia sp |
einfach |
(PCR-Produkt: 246 bp)
RICK-1: 5´-TTCTCAATTCGGTAAGGGC-3´
RICK-2:
5´-ATATTGACCAGTGCTATTTC-3´ |
56 Grad / 45
Zyklen |
(12) |
|
Bartonella
spec. |
einfach |
PCR-Produkt je nach Art 145 bis 251 bp
Bart-1: 5´-CTCTTTCTTCAGATGATGATCC-3´
Bart-2:
5´-AACCAACTGAGCTACAAGCCCT-3´ |
50 Grad / 45
Zyklen |
(15) |
|
HTLV-1 |
|
1. PCR
AV45: 5´-GGACGCGTTRTCRGCTC-3´
AV46: 5´-KGGRGATAGYTGGTAKAGGTA-3´
2. PCR
(PCR-Produkt 100 bp)
AV80:
5´-GGTCTGGAAAAGACAGGGTTG-3´
AV49: 5´-CCCTCCTTCCTCCAGGCCAT-3´ |
50 Grad / 45 Zyklen
50 Grad / 45 Zyklen
|
(17) |
|
Orf Virus |
einfach |
(PCR-Produkt 170 bp)
Orf-F:
5´-TACCAGCCAGAGGATCTCC-3´
Orf-R: 5´-CCACGTCGATAAAGTGTTTGA-3´ |
65°C / 45 Zyklen |
(18) |
|
t(14;18) |
zwei Mal einfach
mbr3/JH
mbr4/JH
|
Länge je
nach Bruchpunkt variabel
mbr3:
5´-TTTGACCTTTAGAGAGTTGCTTTTACG-3´
JH: 5´-acctgaggagacggtgacc-3´
mbr4:
5´-GCAACAGAGAACCATCCCT-3´
JH: 5´-acctgaggagacggtgacc-3´ |
mbr3/JH
55°C / 45 Zyklen
mbr4/JH
58°C / 45 Zyklen |
(19) |
|
Staphylokokken |
einfach |
icaD-F: 5´-ATGGTCAAGCCCAGACAGAG-3´
icaD-R:
5´-CGTGTTTTCAACATTTAATGCAA-3´ |
Touchdown
von 62 Grad nach 52 Grad bei
52°C / 35
Zyklen |
(20) |
|
Streptokokken |
einfach |
(PCR-Produkt 256 bp)
SPEB-1:
5´-GTCAACATGCAGCTACAGGA-3´
SPEB-2:
5´-AATACCAACATCAGCCATCA-3´ |
Touchdown
von 62 Grad nach 52 Grad bei
52°C / 35
Zyklen |
(21) |
|
Pseudomonas spec. |
einfach |
(PCR-Produkt 249 bp)
PS-1: 5´-ATGAACAACGTTCTGAAATTCTCTGCT-3´
PS-2:
5´-CTTGCGGCTGGCTTTTTCCAG-3´ |
52 Grad / 45
Zyklen |
(22) |
Literatur
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rRNA Gene Organization in the Lyme Disease Spirochete, Borrelia burgdorferi.
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Zoechling, N. et al. 1997
Molecular Detection of Treponema pallidum in Secondary and Tertiary
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British Journal of Dermatology 136; 683-86
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Klemen, H. et al. 1998
Multiplex polymerase chain reaction for rapid detection of atypical mycobacteria
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Diagn Mol Pathol 7; 310-16.
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Nishimura, M. 1994
Methods in Pathology. An Improved Method for DNA Diagnosis of Leprosy Using
Formaldehyde-Fixed, Paraffin-Embedded Skin Biopsies.
Modern Pathology 7(2); 253-56
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Evander, M. et al. 1992
Comparison of a One-Step and a Two-Step Polymerase Chain Reaction with
Degenerate General Primers in a Population-Based Study of Human Papillomavirus
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Low Incidence of Epstein-Barr Virus Presence in Primary Cutaneous T-Cell
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Schlupen, E.M. et al. 1995
Molecular Evidence for the Existence of Disseminated Zoster as a Distinct Entity
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Aractingi, S. et al. 1996
Immunohistochemical and Virological Study of Skin in the Papular-Purpuric Gloves
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British Journal of Dermatology 135(4); 599-602
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Bhattacharyya, R. et al. 1996
Development of a Genus Specific Primer Set for Detection of Leishmania
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Tange et al. 1994
Detection of causative agent of spotted fever group rickettsiosis in Japan from
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Melchers, W. et al. 1993
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zurück zur
Tabelle
zum
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