Evaluation of antibody level against Fusobacterium nucleatum in the serological diagnosis of colorectal cancer - PubMed (original) (raw)

Evaluation of antibody level against Fusobacterium nucleatum in the serological diagnosis of colorectal cancer

Hai-Fang Wang et al. Sci Rep. 2016.

Abstract

Fusobacterium nucleatum (F. nucleatum, Fn) is associated with the colorectal cancer (CRC). Fn-infection could induce significant levels of serum Fn-specific antibodies in human and mice. The objective of this study was to identify Fn-infection that elicit a humoral response in patients with CRC and evaluate the diagnostic performance of serum anti-Fn antibodies. In this work, we showed the mean absorbance value of anti-Fn-IgA and -IgG in the CRC group were significantly higher than those in the benign colon disease group and healthy control group (P < 0.001). The sensitivity and specificity of ELISA for the detection of anti-Fn-IgA were 36.43% and 92.71% based on the optimal cut-off. The combination of anti-Fn-IgA and carcino-embryonic antigen (CEA) was better for diagnosing CRC (Sen: 53.10%, Spe: 96.41%; AUC = 0.848). Furthermore, combining anti-Fn-IgA with CEA and carbohydrate antigen 19-9 (CA19-9) (Sen: 40.00%, Spe: 94.22%; AUC = 0.743) had the better ability to classify CRC patients with stages I-II. These results suggested that Fn-infection elicited high level of serum anti-Fn antibodies in CRC patients, and serum anti-Fn-IgA level may be a potential diagnosing biomarker for CRC. Serum anti-Fn-IgA in combination with CEA and CA19-9 increases the sensitivity of detecting early CRC.

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Figures

Figure 1

Figure 1. Detection of the specific antigen of Fn that leads to an intense immune response of CRC patients.

(A) PCR was detected by PCR in stool of 6 CRC patients and healthy controls respectively (C: CRC patients; H: healthy controls; neg: negative control). (B,C) Antigens reactive with anti-Fn-IgA (B) and anti-Fn-IgG (C) were determined by western blotting through incubating with a reference serum dilution of 6 Fn-positive CRC patients or 6 Fn-negative healthy individual as primary antibody. (D) The whole proteins of Fn were separated by 10% SDS-PAGE and then stained with Coomassie brilliant blue (lane 1) and the specific antigens that caused high levels of anti-Fn-IgA were detected by western blotting through incubating with mixed serum samples of 6 CRC patients as primary antibody (lane 2). Of note, Fig. 1B–D were cropped from a single image on the dashed lines to be better presented in the article’s context. The gels have been run under the same conditions and subsequently processed with the same set of materials. These three complete figures could be found respectively in the Supplementary Figs S1 and S2.

Figure 2

Figure 2. Comparison of OD values of anti-Fn-IgA or anti-Fn-IgG in sera from healthy adult human subjects (HS: healthy subjects, n = 200), benign colon disease (n = 150), stage I-II of CRC (n = 55), stage III-IV of CRC (n = 203), the total of CRC patients (n = 258) were individually assayed.

Symbols indicate individual OD value; horizontal lines indicate mean values ± SD. Differences between the five groups were analyzed by Kruskal-Wallis test. (A) anti-Fn-IgA. (B) anti-Fn-IgG.

Figure 3

Figure 3. Diagnostic outcomes for serum anti-Fn-IgA, anti-Fn-IgG, CEA or CA19-9 alone or in combination in the diagnosis of CRC.

(A) ROC curves for the diagnostic strength to identify CRC using anti-Fn-IgA, anti-Fn-IgG, CEA or CA19-9 level. (anti-Fn-IgA: AUC = 0.704; anti-Fn-IgG: AUC = 0.645; CEA: AUC = 0.796; CA19-9: AUC = 0.635). (B) ROC curves for the diagnostic strength to identify CRC using anti-Fn-IgA, CEA or CA19-9. (CEA+ CA19-9: AUC = 0.809; anti-Fn-IgA + CEA+ CA19-9: AUC = 0.858).

Figure 4

Figure 4. Diagnostic outcomes for serum anti-Fn-IgA, anti-Fn-IgG, CEA or CA19-9 alone or in combination in the diagnosis of early stage CRC.

(A) ROC curves for the diagnostic strength to identify early CRC using anti-Fn-IgA, anti-Fn-IgG, CEA or CA19-9 level. (anti-Fn-IgA: AUC = 0.709 anti-Fn-IgG: AUC = 0.612; CEA: AUC = 0.624; CA19-9: AUC = 0.492). (B) ROC curves for the diagnostic strength to identify CRC using anti-Fn-IgA, CEA or CA19-9. (CEA+ CA19-9: AUC = 0.605; anti-Fn-IgA + CEA+ CA19-9: AUC = 0.743).

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References

    1. Gao Z., Guo B., Gao R., Zhu Q. & Qin H.. Microbiota disbiosis is associated with colorectal cancer. Front Microbiol. 6, 20 (2015). - PMC - PubMed
    1. Allen-Vercoe E. & Jobin C.. Fusobacterium and Enterobacteriaceae: Important players for CRC? Immunol Lett. 162, 54–61 (2014). - PMC - PubMed
    1. Keku T. O., McCoy A. N. & Azcarate-Peril A. M.. Fusobacterium sp. and colorectal cancer— cause or consequence? Trends Microbiol. 21, 10.1016/j.tim.2013.08.004 (2013). - DOI - PMC - PubMed
    1. Kostic A. D. et al.. Genomic analysis identifies association of Fusobacterium with colorectal carcinoma. Genome Res. 22, 292–298 (2012). - PMC - PubMed
    1. Castellarin M. et al.. Fusobacterium nucleatum infection is prevalent in human colorectal carcinoma. Genome Res. 22, 299–306 (2012). - PMC - PubMed

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