Molecular Markers for Detecting Schistosoma Species by Loop-Mediated Isothermal Amplification
Table 1
Primer sets selected in this work for amplifying the S. bovis NADH subunit 1 and the ITS-1 region from the genus Schistosoma.
(a) LAMP primers for the S. bovis sequence (LAMP Designer)
NADH subunit 1
Primer
5 pos
3 pos
Length (bp)
Tm (°C)
GC ratio (%)
Sequence
SbF3
219
236
18
56.8
38.9
TTCATTGTTAGGTTGCGT
SbB3
642
619
24
57
33.3
TCTATATTCTACTCTAATCCCTCT
SbFIP
48
TCAGTATCATCTCAAACATCACACTAGTAGTATGTTCTGTCTTAAGTT
SbBIP
45
TTTGTAGTACCTCTGGTTTACATCATTCACTCTCAGACTCTACAT
SbF2
327
349
23
56.8
30.4
AGTAGTATGTTCTGTCTTAAGTT
SbF1c
424
448
25
62.1
36
TCAGTATCATCTCAAACATCACACT
SbB2
593
612
20
57
40
TTCACTCTCAGACTCTACAT
SbB1c
520
544
25
61.8
36
TTTGTAGTACCTCTGGTTTACATCA
SbLF
364
388
25
61.9
36
ACTTAGACCATGAACATCAACCTAT
SbLB
560
584
25
61.9
40
TACTAAGTGAGAGTAATCGAACACC
(b) LAMP primers for the genus Schistosoma sequence (Primer Explorer V5)
ITS-1
Primer
5 pos
3 pos
Length (bp)
Tm (°C)
GC ratio (%)
Sequence
SF3
2
19
18
59.7
61
TTGACCGGGGTACCTAGC
SB3
200
218
19
59.5
53
CGTGAATGGCAAGCCAAAC
SFIP
39
ATCGCCCTTGGCAGATCAGGCTGTCGTATGCCCTGATGG
SBIP
40
ATATGCATGCAAATCCGCCCCGCGGATCGCTTCAACAGTGTA
SF2
20
38
19
59.2
58
CTGTCGTATGCCCTGATGG
SF1c
61
80
20
64.2
60
ATCGCCCTTGGCAGATCAGG
SB2
180
199
20
59.5
50
CGGATCGCTTCAACAGTGTA
SB1c
135
156
20
65.9
55
ATATGCATGCAAATCCGCCCCG
SLF
39
60
22
60.4
45
CAGATCAGGCAACCCGAAAG
For S. bovis (Sb) and genus Schistosoma (S): F3=forward outer primer; B3=backward outer primer; FIP=forward inner primer (comprising F1c and F2 sequences); BIP=backward inner primer (comprising B1c and B2 sequences); LF=loop forward primer; LB=loop backward primer.
