System for diagnosing hormonal femoral head necrosis through molecular markers
阅读说明：本技术 通过分子标志物诊断激素性股骨头坏死的系统 (System for diagnosing hormonal femoral head necrosis through molecular markers ) 是由 陈卫衡 张彦琼 林娜 李泰贤 王荣田 于 2019-02-03 设计创作，主要内容包括：本公开涉及一种通过分子标志物诊断激素性股骨头坏死的系统,其中,所述分子标志物为BIRC3、CBL、CCR5、LYN、PAK1、PTEN、RAF1和TLR4。本公开所提供的系统和用途可以应用于通过检测分子标志物的表达量对SONFH高危人群作出评估和诊断,并协助作出较早期诊断,为指导临床诊疗提供有效依据。(The present disclosure relates to a system for diagnosing hormonal femoral head necrosis by molecular markers, wherein the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4. The system and the application provided by the disclosure can be applied to assessment and diagnosis of SONFH high risk groups by detecting the expression quantity of the molecular marker, and assist in making early diagnosis, so as to provide an effective basis for guiding clinical diagnosis and treatment.)
1. A system for diagnosing hormonal femoral head necrosis is characterized by comprising a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of a molecular marker of an individual patient with femoral head necrosis; wherein the molecular marker is BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4; the computing device comprises a memory and a processor; the memory has a computer program stored therein, and the processor is configured to execute the computer program stored in the memory to implement a modeling algorithm and an algorithm of a discriminant function as shown in equation (1); the modeling algorithm is a minimum deviation two-times algorithm;
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+f8(c8)-b]the compound of the formula (1),
in the formula (1), f (c) represents the diagnosis result of the hormonal femoral head necrosis, f (c) has a return value of 1 indicating that the hormonal femoral head necrosis is established, and has a return value of-1 indicating that the hormonal femoral head necrosis is not established; c. C1～c8In turn, the relative expression of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR4, respectivelyAn amount; the relative expression amount refers to the ratio of the expression amount relative to an internal reference; f. of1(c1)～f8(c8) The kernel functions are obtained by training according to the modeling algorithm, and b is the critical score values obtained by training according to the modeling algorithm.
2. The system according to claim 1, wherein the system further comprises a means for detecting the expression level of the molecular marker; the detection device comprises a molecular marker expression quantity detection chip and a chip signal reader, wherein the molecular marker expression quantity detection chip comprises probes for respectively detecting the expression quantities of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
3. The system of claim 2, wherein the chip for detecting the expression level of the molecular marker further comprises an internal reference probe, and the internal reference probe is a probe for detecting the expression level of GAPDH or β -Actin.
4. The system according to claim 2, wherein the detection device comprises a real-time quantitative PCR instrument and real-time quantitative PCR primers of the molecular markers, and the real-time quantitative PCR primers of the molecular markers comprise real-time quantitative PCR primers for respectively detecting the expression levels of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
5. The system of claim 4, wherein the real-time quantitative PCR primers of the molecular marker further comprise an internal reference primer, and the internal reference primer is a real-time quantitative PCR primer for detecting GAPDH or beta-Actin.
6. The system of claim 1, wherein in formula (1), the internal reference is GAPDH, f1(c1)＝-0.097×c1，f2(c2)＝0.019×c2，f3(c3)＝-0.316×c3，f4(c4)＝0.292×c4，f5(c5)＝0.681×c5，f6(c6)＝0.375×c6，f7(c7)＝0.428×c7，f8(c8)＝0.129×c8B is 0.040; alternatively, the first and second electrodes may be,
the internal reference is β -Actin, f1(c1)＝-0.012×c1，f2(c2)＝-0.329×c2，f3(c3)＝-0.254×c3，f4(c4)＝0.332×c4，f5(c5)＝0.561×c5，f6(c6)＝0.422×c6，f7(c7)＝0.469×c7，f8(c8)＝-0.052×c8，b＝0.024。
7. Use of a reagent for quantitatively detecting molecular markers in the preparation of a product for diagnosing hormonal femoral head necrosis, wherein the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
8. Use according to claim 7, wherein the quantitative detection of molecular markers is carried out by:
1) obtaining a serum sample of a patient with femoral head necrosis;
2) determining the expression level of the molecular marker in the serum sample.
9. A kit for diagnosing hormonal femoral head necrosis, wherein the kit comprises a reagent for quantitatively detecting molecular markers, and the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
10. A molecular marker combination for diagnosing hormonal femoral head necrosis, wherein the molecular marker combination comprises BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Technical Field
The present disclosure relates to the field of clinical testing techniques, and in particular, to a system for diagnosing hormonal femoral head necrosis by molecular markers.
Background
Femoral Head necrosis (Osteoneprosis of the Femoral Head, ONFH) is a common disease of orthopedics, the number of the disease attack in China currently exceeds ten million, and 15-30 ten thousand new patients are newly added every year. The Femoral Head necrosis with different causes can be divided into traumatic Femoral Head necrosis (non-traumatic Osteonecrosis of the bone of the femur neck, Femoral Head dislocation and other traumatic factors) and non-traumatic Femoral Head necrosis (NONFH), the former is caused by the traumatic factors such as Femoral neck fracture and Femoral Head dislocation, and the latter is caused by the reasons of glucocorticoid drug treatment, alcohol addiction and the like.
Hormone-induced femoral head necrosis (SONFH) is a non-traumatic femoral head necrosis caused by long-term hormone use, and the incidence rate of the SONFH exceeds that of femoral head necrosis caused by trauma at present.
At present, the most effective method for diagnosing the femoral head necrosis is MRI examination, the early diagnosis rate reaches more than 90 percent, but the further development of the necrosis is difficult to prevent after the diagnosis is confirmed. Therefore, for the high risk population of SONFH, it is very important to predict the occurrence of osteonecrosis and to prevent it in advance.
Disclosure of Invention
The purpose of the present disclosure is to provide a kit and a system for diagnosing hormonal femoral head necrosis by using a molecular marker, and the use of the molecular marker in diagnosing the hormonal femoral head necrosis.
To achieve the above object, a first aspect of the present disclosure: the system comprises a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of the molecular marker of an individual patient with femoral head necrosis, and the output device is used for outputting the diagnosis result of the femoral head necrosis; wherein the molecular marker is BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4; the computing device comprises a memory and a processor; the memory has a computer program stored therein, and the processor is configured to execute the computer program stored in the memory to implement a modeling algorithm and an algorithm of a discriminant function as shown in equation (1); the modeling algorithm is a minimum deviation two-times algorithm;
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+f8(c8)-b]in the formula (1), f (c) represents a diagnosis result of the hormonal femoral head necrosis, f (c) has a return value of 1 indicating that the hormonal femoral head necrosis is established, and has a return value of-1 indicating that the hormonal femoral head necrosis is not established; c. C1～c8Sequentially and respectively representing the relative expression amounts of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4; the relative expression amount refers to the ratio of the expression amount relative to an internal reference; f. of1(c1)～f8(c8) The kernel functions are obtained by training according to the modeling algorithm, and b is the critical score values obtained by training according to the modeling algorithm.
Optionally, the system further comprises a device for detecting the expression level of the molecular marker; the detection device comprises a molecular marker expression quantity detection chip and a chip signal reader, wherein the molecular marker expression quantity detection chip comprises probes for respectively detecting the expression quantities of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Optionally, the chip for detecting the expression level of the molecular marker further comprises an internal reference probe, wherein the internal reference probe is a probe for detecting the expression level of GAPDH or β -Actin.
Optionally, the detection device comprises a real-time quantitative PCR instrument and real-time quantitative PCR primers of the molecular markers, wherein the real-time quantitative PCR primers of the molecular markers comprise real-time quantitative PCR primers for respectively detecting the expression levels of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Optionally, the real-time quantitative PCR primers of the molecular marker further include an internal reference primer, and the internal reference primer is a real-time quantitative PCR primer for detecting GAPDH or β -Actin.
Optionally, in formula (1), the internal reference is GAPDH, f1(c1)＝-0.097×c1，f2(c2)＝0.019×c2，f3(c3)＝-0.316×c3，f4(c4)＝0.292×c4，f5(c5)＝0.681×c5，f6(c6)＝0.375×c6，f7(c7)＝0.428×c7，f8(c8)＝0.129×c8B is 0.040; alternatively, the first and second electrodes may be,
the internal reference is β -Actin, f1(c1)＝-0.012×c1，f2(c2)＝-0.329×c2，f3(c3)＝-0.254×c3，f4(c4)＝0.332×c4，f5(c5)＝0.561×c5，f6(c6)＝0.422×c6，f7(c7)＝0.469×c7，f8(c8)＝-0.052×c8，b＝0.024。
In a second aspect of the present disclosure: the application of a reagent for quantitatively detecting molecular markers in preparing a product for diagnosing the hormonal femoral head necrosis is provided, wherein the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Alternatively, the quantitative detection of the molecular marker is performed by:
1) obtaining a serum sample of a patient with femoral head necrosis;
2) determining the expression level of the molecular marker in the serum sample.
A third aspect of the disclosure: provided is a kit for diagnosing hormonal femoral head necrosis, wherein the kit comprises a reagent for quantitatively detecting molecular markers, and the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
A fourth aspect of the present disclosure: provided is a molecular marker combination for diagnosing hormonal femoral head necrosis, wherein the molecular marker combination comprises BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Through the technical scheme, the system and the application provided by the disclosure can be applied to assessment and diagnosis of SONFH high-risk people by detecting the expression quantity of the molecular marker, and assist in making early diagnosis, so that an effective basis is provided for guiding clinical diagnosis and treatment.
Additional features and advantages of the disclosure will be set forth in the detailed description which follows.
Drawings
The accompanying drawings, which are included to provide a further understanding of the disclosure and are incorporated in and constitute a part of this specification, illustrate embodiments of the disclosure and together with the description serve to explain the disclosure without limiting the disclosure. In the drawings:
fig. 1-2 are ROC graphs obtained by diagnosing hormonal femoral head necrosis using the molecular markers of the present disclosure in examples, wherein fig. 1 is internally referenced as GAPDH, and fig. 2 is internally referenced as β -Actin.
Detailed Description
The following detailed description of specific embodiments of the present disclosure is provided in connection with the accompanying drawings. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.
The first aspect of the disclosure: the system comprises a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of the molecular marker of an individual patient with femoral head necrosis, and the output device is used for outputting the diagnosis result of the femoral head necrosis; wherein the molecular marker is BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4; the computing device comprises a memory and a processor; the memory has a computer program stored therein, and the processor is configured to execute the computer program stored in the memory to implement a modeling algorithm and an algorithm of a discriminant function as shown in equation (1); the modeling algorithm is a minimum deviation two-times algorithm;
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+f8(c8)-b]the compound of the formula (1),
in the formula (1), F (c) represents the diagnosis result of the hormonal femoral head necrosis, and F (c) returns of 1 represent the hormoneWhen the return value is-1, the hormonal femoral head necrosis is not established; c. C1～c8Sequentially and respectively representing the relative expression amounts of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4; the relative expression amount refers to the ratio of the expression amount relative to an internal reference; f. of1(c1)～f8(c8) The kernel functions are obtained by training according to the modeling algorithm, and b is the critical score values obtained by training according to the modeling algorithm. The least-squares multiplication algorithm and the operation and training method thereof are known conventional methods.
In accordance with the present disclosure, BIRC3 is referenced 330 in the NCBI database; CBL reference number 867 in NCBI database; CCR5 reference number 1234 in the NCBI database; LYN is referenced 4067 in NCBI database; PAK1 reference number 5058 in NCBI database; reference number of PTEN in NCBI database is 5728; reference number 5894 of RAF1 in NCBI database; TLR4 is referenced 7099 in the NCBI database.
According to the present disclosure, the system may further comprise a device for detecting the expression level of the molecular marker. The detection device comprises a molecular marker expression quantity detection chip and a chip signal reader, wherein the molecular marker expression quantity detection chip comprises probes for respectively detecting the expression quantities of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4. Or the detection device comprises a real-time quantitative PCR instrument and real-time quantitative PCR primers of the molecular markers, and the real-time quantitative PCR primers of the molecular markers comprise real-time quantitative PCR primers for respectively detecting the expression levels of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
According to the disclosure, the chip for detecting the expression level of the molecular marker further comprises an internal reference probe, wherein the internal reference probe is a probe for detecting the expression level of GAPDH or beta-Actin. The real-time quantitative PCR primer of the molecular marker also comprises an internal reference primer, and the internal reference primer is a real-time quantitative PCR primer for detecting GAPDH or beta-Actin.
According to the present disclosure, when the internal reference is GAPDH, as a set of reference values obtained by training, in formula (1), f1(c1)＝-0.097×c1，f2(c2)＝0.019×c2，f3(c3)＝-0.316×c3，f4(c4)＝0.292×c4，f5(c5)＝0.681×c5，f6(c6)＝0.375×c6，f7(c7)＝0.428×c7，f8(c8)＝0.129×c8B is 0.040; that is, the discriminant function can be simplified as shown in equation (2):
F(c)＝sgn[-0.097×c1+0.019×c2-0.316×c3+0.292×c4+0.681×c5+0.375×c6+0.428×c7+0.129×c8-0.040]formula (2).
Or when the internal parameter is β -Actin, the internal parameter is used as a group of reference values obtained by training, and in the formula (1), f1(c1)＝-0.012×c1，f2(c2)＝-0.329×c2，f3(c3)＝-0.254×c3，f4(c4)＝0.332×c4，f5(c5)＝0.561×c5，f6(c6)＝0.422×c6，f7(c7)＝0.469×c7，f8(c8)＝-0.052×c8B is 0.024; that is, the discriminant function can be simplified as shown in equation (3):
F(c)＝sgn[-0.012×c1-0.329×c2-0.254×c3+0.332×c4+0.561×c5+0.422×c6+0.469×c7-0.052×c8-0.024]formula (3).
In addition, f is1(c1)～f8(c8) And b may vary depending on the bias of the means for detecting the expression level of the molecular marker, or may vary depending on factors such as the size of the data scale of the training data set. The discriminant functions shown in equations (2) and (3) are obtained by training the inventors of the present disclosure with a modeling algorithm of least-squares multiplication according to the data in the examples, and do not limit the scope of the present disclosure.
In a second aspect of the present disclosure: the application of a reagent for quantitatively detecting molecular markers in preparing a product for diagnosing the hormonal femoral head necrosis is provided, wherein the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Further, the quantitative detection of the molecular marker is performed by the following steps:
1) obtaining a serum sample of a patient with femoral head necrosis;
2) determining the expression level of the molecular marker in the serum sample.
Wherein, the method of step 2) may be a qPCR method.
The system and the application provided by the disclosure can be applied to assessment and diagnosis of SONFH high risk groups by detecting the expression quantity of the molecular marker, and assist in making early diagnosis, so as to provide an effective basis for guiding clinical diagnosis and treatment.
A third aspect of the disclosure: provided is a kit for diagnosing hormonal femoral head necrosis, wherein the kit comprises a reagent for quantitatively detecting molecular markers, and the molecular markers are BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
Further, the reagents may be qPCR reagents.
A fourth aspect of the present disclosure: provided is a molecular marker combination for diagnosing hormonal femoral head necrosis, wherein the molecular marker combination comprises BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
In a particularly preferred embodiment of the present disclosure, the molecular marker combination consists of BIRC3, CBL, CCR5, LYN, PAK1, PTEN, RAF1 and TLR 4.
The kit and the molecular marker combination provided by the disclosure can be applied to assessment and diagnosis of SONFH high-risk people by detecting the expression quantity of the molecular marker, assist in making early diagnosis and provide effective basis for guiding clinical diagnosis and treatment.
The present disclosure will be described in detail below with reference to examples. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.