A new type of Ag nanofilm with high efficiency is prepared and used here, acting as a near-infrared surface-enhanced Raman spectroscopy (NIR-SERS) substrate to detect oxyhemoglobin (OxyHb) for 22 cervical cancer patients and 22 healthy females. NIR-SERS results show that there is striking spectral difference of the OxyHb between healthy females and cervical cancer patients. Meanwhile, principal component analysis (PCA) combined with independent sample T-test analysis is employed to analyze the measured NIR-SERS spectra, and it is found that the measured NIR-SERS spectra of the two groups are separated into two distinct clusters in terms of the sensitivity of 86.4% and the specificity of 86.4%. Tentative assignments of the Raman bands of the measured NIR-SERS spectra are also performed, and the results suggest that cancer specific changes at molecular level, including pyrrole ring and vibration mode of the OxyHb molecules are significant. The NIR-SERS detection of OxyHb for cervical cancer patients based on PCA combined with independent sample T-test is expected to develop into a new diagnostic tool for cervical cancer.