Method for establishing indications for prostate biopsy

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to urology. To establish the indications for prostate biopsy, prostate secretion is analysed. Kallikrein (KK) activity (mcmol x min-1 x l-1) and angiotensin converting enzyme (ACE) activity (mcmol x min-1 x l-1) are evaluated. Provided 18.20≤KK≤58.62 and 32.16≤ACE≤51.20, prostate biopsy is considered to be indicated.

EFFECT: method provides higher reliability of establishing the indications for prostate biopsy, is easy to implement, and does not require considerable material costs.

1 ex

 

The invention relates to medicine, namely to urology, and will be used to determine the indications for biopsy of the prostate.

Development of methods for determining the indications for biopsy of the prostate gland, suitable for mass screening, is one of the main tasks of modern urology.

In the United States every year, which amounts to 300 thousand new cases of prostate cancer (PC). 30 thousand men per year in the US die from this disease. The problem of determining the indications for prostate biopsy is extremely relevant and in Russia, where every year, which amounts to about 20,000 new cases of PC. The incidence is 25.6 per 100 thousand population. An annual growth rate of 63.9% (Bealestreet, Kamniska. Hormonal therapy of patients with prostate cancer. J. "Oncourology", 2007-2008, M., special issue, pp.2-5).

There are many ways to define the indications for prostate biopsy. The modern way to define the indications for prostate biopsy based on the joint application definition prostatespecific antigen (PSA) and digital rectal exam (IF), in conjunction with transrectal ultrasound (transrectal us) prostate. The combination of these techniques biopsy of the prostate is shown in 80% of patients. But overall RUSNANO, that DOG standard, prostatespecific the marker, interspecifically to 20% (Pannek J., Partin A.W., Prostate-specific antigen: whats new in 1997. Oncology (Williston Park) 1997; 11: 1273-82). Diagnostic value indicators such as the ratio of free PSA to total, the rate of increase of PSA, density and range of normative age-related PSA values, is under study.

Sensitivity is not higher than 50%. Many urologists find this research is justified only for technical support biopsy.

Prostate biopsy is the only reliable tool for the detection of an existing prostate cancer (Chibichan MB, Kogan M.I. Phenomenon of migration stage prostate cancer after radical prostatectomy. J. "Bulletin of the southern scientific center, Rostov-on-don, 2009, Volume 5, No. 1, p.11-17). However, even with 10-12 point biopsy of the prostate 10-35% of men with negative initial biopsy have cancer at repeat biopsy. Urologists have to repeat uncomfortable for patients with prostate biopsy, accompanied by 25% of patients with severe pain, hematospermia, hematuria (Djavan B, et al. J Urol 2000; 163:1144-9; Roehl KA, et al. J Urol 2002; 167:2435-9); Heidenreich A, et al. EurUrol 2008; 53:68-80 (EAU guidelines on prostate cancer 2008).

Although biopsy remains the "gold standard" for detection of prostate cancer VC is SHL, for making decisions about the need for biopsy requires the development of new methods for determining the indications for biopsy of the prostate.

Conducted research on the medical-scientific and patent literature found different ways to determine the indications for biopsy of the prostate.

The study I.Rehman, A.R.Azzouzi, J.W.F.Catto, S.Alien, S.S.Cross, K.Feely, M.Meuth, F.C.Hamdy Proteomic analysis of voided urine after prostatic massage from patients with prostate cancer: a pilot study. Urology 64:1238-1243, 2004, describes how to determine the indications for biopsy of the prostate gland, where patients with suspected FPW investigated urine after massage of the prostate gland. Urine was centrifuged to remove cells and debris, spent separation and protein markers by two-dimensional polyacrylamide gel electrophoresis and identified protein markers. Detection in urine protein Calgranulin B/MRP-14 biopsy of the prostate was considered shown.

In the same study described another way to define the indications for prostate biopsy, where a similar method was investigated urine of the patient, and in the presence of protein in it Transferrin A chain (36 kDa) biopsy of the prostate was considered shown.

The disadvantages of the above methods is the low specificity of the markers, the complexity of the technological platforms: the need for special equipment the equipment and staff of specially trained staff, lack of reliability.

The closest technical solution adopted by the us as a prototype, is how to define the indications for prostate biopsy described Bussemakers MJG, et al. (Cancer Res 1999; 59:5975-9), US Patent No. 5185439 at which conduct the study of the biological material in the third portion of urine after massage of the prostate gland. The method is based on the technology of nucleic acid amplification, used for detecting mRNA Prostate specific gene 3 (Prostate Cancer Gene 3 (RS)) in the urine samples of men. Urine is treated by adding a Transport Medium for Urine (SCI), which is the lysis of cells and stabilization of RNA. Next, the analysis counts RSA and PSA mRNA ratio RA/PSA mRNA define the index RSA. When the index PCA 3 more 100 - biopsy of the prostate is considered shown.

The disadvantage of this method of determining the indications for prostate biopsy is its complexity, high cost of research, which hampers the possibility of large-scale implementation in practice, the lack of accuracy is 89%.

The aim of the present invention is to simplify the method of determining the indications for biopsy of the prostate, reducing the cost of its implementation, improving the reliability determination by the Azania biopsy of the prostate.

The objective is achieved by determining in a biological fluid of the patient - the secret of the prostate gland activity (µmol × min-1× l-1) kallikrein (KK) and activity (µmol × min-1× l-1) angiotensin-converting enzyme (ACE)inhibitors. Under the conditions 18,20≤QC≤58,62 and 32,16≤ACE≤51,20 biopsy of the prostate is considered shown.

The proposed method of determining the indications for prostate biopsy is as follows.

The patient after the massage the prostate gland taking the secret of the prostate gland, which determine the activity (µmol × min-1× l-1) kallikrein (KK) and activity (µmol × min-1× l1) angiotensin-converting enzyme (ACE)inhibitors.

Before the study the secret of the prostate gland is diluted in the ratio of 1:9 with saline to a volume of at least 0.5 ml

The activity of kallikrein determined by a spectrophotometric method for rate of hydrolysis of N-benzoyl-1-arginine ethyl ester (BAEE) (Pashina T.S., Krinsky AV Simplified method for the determination of kallikrein and kallikrein serum (plasma) blood in norm and in certain pathological conditions. Questions of medical chemistry. 1974; 20(6): 660-663).

The activity of kallikrein is expressed in (µmol × min-1× l -1).

The activity of angiotensin-converting enzyme (ACE) is determined by a spectrophotometric method using a synthetic substrate ACE forilalililla-glycylglycine (BASSOON) (Golikov P.P., Nikolaev POSTGRADUATE rapid method for determining the activity of angiotensin-converting enzyme in serum // Klin. Lab. Diagnosis. - 1998. - 1. - C.11-13).

Enzyme activity is expressed as µmol × min-1× l-1.

Practical feasibility of the proposed method of determining the indications for prostate biopsy confirmed an example from clinical practice.

Example

The patient badgers S.L., 59 years old, was admitted to outpatient evaluation at the clinic of urology of the Rostov Gosmeduniversiteta (Rostov state medical University) with a preliminary diagnosis of prostatic hyperplasia.

Complaints: frequent and painful, difficulty urinating. The history of the disease: these complaints symptoms occur within 2 years.

The results has shown that the boundaries of the prostate fuzzy; iron is moderately enlarged, hardened, non-uniform.

Evaluation of urinary international scale assessment of prostatic symptoms (IPPS) - 18 points.

Urinalysis is normal. Laboratory diagnosis: PSA=9,426 ng/ml. the Results of transrectal us prostate cancer: volume issue the ATA - 26,3 cm3; residual urine volume - 70 cm3.

To determine the indications for prostate biopsy the patient study was conducted according to the claimed method.

The patient after a massage of the prostate was carried out by the fence secret of the prostate gland, which determined the activity concentration of kallikrein (KK) and the activity of angiotensin-converting enzyme (ACE).

Before the study obtained the secret of a prostate cancer patient was diluted in the ratio of 1:9 with saline to a volume of 0.5 ml.

The activity of kallikrein (KK) was determined spectrophotometrically by the method of the speed of hydrolysis of N-benzoyl-1-arginine ethyl ester (BAEE) (Pashina T.S., Krinsky AV Simplified method for the determination of kallikrein and kallikrein serum (plasma) blood in norm and in certain pathological conditions. Questions of medical chemistry 1974; 20(6):660-663).

The activity of kallikrein was expressed in µmol × min-1× l-1. It was obtained the value of QC=34,48 mmol × min-1× l-1.

The activity of angiotensin-converting enzyme (ACE) inhibitors was determined by a spectrophotometric method using a synthetic substrate ACE forilalililla-glycylglycine (FAGG) (Golikov P.P., Nikolaev POSTGRADUATE rapid method for determining the activity of any antiprogressive enzyme in serum // Klin. Lab. Diagnosis. - 1998. - 1. - C.11-13).

Enzyme activity was expressed in mmol × min-1× l-1. It was obtained the value of PFA=50,14 mmol × min-1× l-1.

Because they were made conditions

18,20≤QC=34,48≤58,62 and

32,16≤PFA=50,14≤51,20,

a prostate biopsy was considered shown.

The patient was performed a biopsy of the prostate. The results morfologicheskogo studies of biopsy, the patient was diagnosed with adenocarcinoma of the prostate T1cN0M0.

The diagnosis is confirmed the feasibility and need for the given patient biopsy of the prostate.

In the clinic of urology of Rostov state medical University us study was conducted according to the claimed method 20 outpatients with suspected prostate cancer.

All patients received values of QC activity and the activity of the enzyme corresponded to the conditions 18,20≤QC≤58,62 and 2.16≤ACE≤51,20, so according to the claimed method biopsy of the prostate to all of them was shown.

19 patients according to morphological studies was diagnosed with prostate cancer. Thus, the accuracy of the proposed method of determination of indications for prostate biopsy was 95%. The cost of research according to the claimed methods which have amounted to two thousand rubles per patient. The proposed method does not require a staff of specially trained personnel, can be performed in an outpatient setting and, therefore, can be widely adopted in clinical practice.

The cost of conducting studies to determine the indications for prostate biopsy according to the prototype is 350-400 euros. The accuracy of the method is 89% (J. Groskopf, S.M.Aubin, J.Mathis, J.Pham, P.Hodge, M.L.Macairan, L.S.Marks, and H.Rittenhouse. 2006. APTIMA PCA3 Molecular Urine Test: Development of a Method to Aid in the Diagnosis of Prostate Cancer. Clin Chem. 52: 1089-95; Shappel S.B. Clinical utiliti of prostate carcinoma molecular diagnostic tests. Rev Urol. 2008; 10(1): 44-69). The method is difficult to implement, is not reliable enough, preventing its widespread implementation.

Thus, compared with the prototype of the proposed method for the determination of indications for prostate biopsy is more reliable, easy to implement and significantly cost-effective.

The method of determining the indications for prostate biopsy involving the study of biological material, characterized in that the secret of the prostate gland determine the activity (µmol · min-1· l-1) kallikrein (KK) and activity (µmol · min-1· l-1) angiotensin-converting enzyme (ACE) inhibitors and subject to the conditions 18,20≤QC≤58,62 and 32,16≤ACE≤51,20 biopsy of the prostate is considered shown.



 

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2 ex

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