Introduction
Methods
Patients and controls
Characteristics | CT (N = 21) | AIS (N = 69) | P value | |||
---|---|---|---|---|---|---|
Male | Female | Male | Female | |||
Case NO | 6 | 15 | 17 | 52 | 0.777 | |
Age (years) | 13.17 ± 1.05 | 14.40 ± 0.75 | 14.47 ± 0.39 | 14.27 ± 0.25 | 0.599a | |
0.590b | ||||||
Major curve Cobb Angle (o) | – | – | 36.76 ± 4.51 | 35.40 ± 2.09 | 0.762 | |
Lenke types | 1 | – | – | 11 | 29 | – |
5 | – | – | 4 | 20 | – | |
2 | – | – | 1 | 1 | – | |
3 | – | – | 0 | 1 | – | |
4 | – | – | 1 | 1 | – | |
6 | – | – | 0 | 0 | – |
Extraction of ccf DNA in plasma
Real time PCR (qPCR)
GENE | Primer sequence (5′–3′) | Length (bp) |
---|---|---|
GAPDH
| F: GGAGAAGCTGAGTCATGGGT | 219 |
R: AAGACGGAATGGGGAGAAGG | ||
ACTB
| F: CTGGAACGGTGAAGGTGACA | 65 |
R: CGGCCACATTGTGAACTTTG | ||
MT-ND1
| F: CCCTAAAACCCGCCACATCT | 214 |
R: CCGATCAGGGCGTAGTTTGA |
Statistical analysis
Results
General physiological features of patients with AIS and the control group
Alteration of plasma ccf n-DNA and ccf mt-DNA levels in AIS patients compared with those in controls
Test | Cutoff (GE/ml) | AUC (95% CI) | Sensitivity | Specificity | P value |
---|---|---|---|---|---|
n-DNA-GAPDH | 732 | 0.589 (0.456–0.722) | 0.551 | 0.667 | 0.217 |
n-DNA-ACTB | 6953 | 0.657 (0.525–0.789) | 0.710 | 0.619 | 0.030 |
mt-DNA | 1,684,972 | 0.562 (0.440–0.683) | 0.319 | 0.905 | 0.393 |
Association of sex with the different plasma ccf n-DNA and ccf mt-DNA levels in AIS patients and controls
Test | Cutoff (GE/ml) | AUC (95% CI) | Sensitivity | Specificity | P value | |
---|---|---|---|---|---|---|
Male | n-DNA-GAPDH | 3170 | 0.667 (0.384–0.950) | 0.706 | 0.667 | 0.234 |
n-DNA-ACTB | 6940 | 0.725 (0.503–0.948) | 0.588 | 0.833 | 0.107 | |
mt-DNA | 2,178,401 | 0.686 (0.443–0.930) | 0.588 | 0.833 | 0.183 | |
Female | n-DNA-GAPDH | 732 | 0.567 (0.417–0.716) | 0.635 | 0.600 | 0.434 |
n-DNA-ACTB | 7291 | 0.636 (0.476–0.796) | 0.769 | 0.533 | 0.111 | |
mt-DNA | 1,684,972 | 0.519 (0.379–0.660) | 0.269 | 0.933 | 0.821 |
Association of the Lenke type with different plasma ccf n-DNA and ccf mt-DNA levels in AIS patients and controls
Test | Cutoff (GE/ml) | AUC (95% CI) | Sensitivity | Specificity | P value | |
---|---|---|---|---|---|---|
Lenke 1 | n-DNA-GAPDH | 729 | 0.586 (0.438–0.733) | 0.667 | 0.550 | 0.274 |
n-DNA-ACTB | 6940 | 0.675 (0.532–0.818) | 0.619 | 0.750 | 0.026 | |
mt-DNA | 4,089,116 | 0.619 (0.477–0.762) | 0.450 | 0.810 | 0.129 | |
Lenke 5 | n-DNA-GAPDH | 732 | 0.629 (0.465–0.793) | 0.625 | 0.667 | 0.139 |
n-DNA-ACTB | 6953 | 0.637 (0.473–0.801) | 0.667 | 0.619 | 0.116 | |
mt-DNA | 164,261,352 | 0.554 (0.376–0.731) | 0.375 | 1.000 | 0.539 |