# Method for express-diagnosis and differential diagnosis of urogenital infection

FIELD: medicine, laboratory diagnosis.

SUBSTANCE: method involves determination of the patient blood content of globulin-alpha 1, globulin-beta, globulin-gamma and the total bilirubin content followed by calculation of diagnosis indices for the patient (Y_{1}, Y_{2}, Y_{3}) by using the computer program "Statistica 1.5" and introducing values X_{1}, X_{2}, X_{3} and X_{4} in computer wherein X_{1} means globulin-alpha 1 value; X_{2} means globulin-beta value; X_{3} means globulin-gamma value; X_{4} means total bilirubin value. Obtained values of diagnosis indices for the individual patient (Y_{1}, Y_{2}, Y_{3}) are compared with average values of diagnosis indices (Y_{1}', Y_{2}', Y_{3}') for different urogenital infections followed by comparison by sign and value. By the maximal coincidence of diagnosis index values for the individual patient with average diagnosis index values urogenital disease is diagnosed and the following diagnosis index average values are used: for chlamydiosis: Y_{1}' = -2; Y_{2}' = -0.1; Y_{3}' = -0.2; for mycoplasmosis: Y_{1}' = 2; Y_{2}' = 0.8; Y_{3}' = -0.04; for ureaplasmosis: Y_{1}' = 2; Y_{2}' = -1; Y_{3}' = 0.02; for health persons: Y_{1}' = -2; Y_{2}' = 0.1; Y_{3}' = 0.2. Invention provides the development of a method for express-diagnosis of infection at initial stage and diagnosis of atypical forms that occur in these diseases, and differential diagnosis of chlamydiosis, mycoplasmosis and ureaplasmosis. Invention can be used for carrying out the differential diagnosis of chlamydiosis, mycoplasmosis and ureaplasmosis.

EFFECT: improved method for express-diagnosis.

2 tbl, 4 ex

The present invention relates to medicine, namely to laboratory diagnosis, and can be used for rapid diagnosis and differential diagnosis of chlamydia, mycoplasmosis and a ureaplasmosis.

The prevalence among the population chlamydia, mycoplasmosis and a ureaplasmosis, their role in the pathogenesis of a number of obstetric and perinatal diseases require to proceed with a more detailed study of this problem [3]. Screening-screening for chlamydial and arenicolsterol infection with subsequent readjustment is one of the reserves reducing morbidity [1]. The effectiveness of existing methods of diagnosis varies depending on the clinical form of infection [5]. Often these diseases are hiding under the guise of various therapeutic pathology and remain undetected, so there is a need for new, more sophisticated methods in the diagnosis of these infections.

In the literature we have not met the ways rapid diagnosis of chlamydia, mycoplasmosis and a ureaplasmosis on biochemical parameters of blood [2].

Traditional, the most appropriate methods of diagnosis of these diseases the following: PCR (polymerase chain reaction), ELISA (enzyme-linked immunosorbent assay), DFA (direct immunofluorescence reaction), bacteriological seeding [4].

PCR-based detection Genet the ical material, which is specific for each species of microorganisms. ELISA is based on the reaction of antigen - antibody by determining the optical density detected antibodies to the pathogen. PIF is also based on the reaction of antigen - antibody, diagnosticum is an antibody to the constituents of the cells of the pathogen, labeled fluorescein. Bacteriological culture of the liquid diagnostic environment based on the ability of the medium during growth of the microorganism to change color due to the manifestation of its enzymatic activity.

Because the ELISA method is proposed invention one common characteristic (the study of blood), this method can be considered the closest to the technical essence.

All these methods have, along with the advantages and significant disadvantages. Typically, they will detect the disease in its active or advanced stage when the patients are already difficult to treat [6]. In addition, IFA, DFA and PCR can give false positive results, and of bacteriological seeding, however, does not always reveal an infection.

Therefore, the object of the invention is to develop a method of rapid diagnosis of infection in its initial stages and diagnosis of atypical forms, which are often found in these diseases, as well as differential diagnosis of chlamydia, mycoplasmosis, ureaplasmosis.

The task in the way of rapid diagnosis and differential diagnosis of urogenital infections is achieved by the fact that in the blood of the subject determine the value of alpha 1 globulin, beta globulin, gamma globulin, and total bilirubin, and then determine the value of the diagnostic indicators of the subject (Y_{1}, Y_{2}, Y_{3}) using the computer program Statistica 1.5”, typing in the computer

X_{1}, X_{2}, X_{3}and X_{4},

where X_{1}- the value of the alpha 1 globulin;

X_{2}the value of beta globulin;

X_{3}the value of gamma globulin;

X_{4}- the value of total bilirubin;

obtained values of the diagnostic indicators for this particular patient (Y_{1}, Y_{2}, Y_{3}) compare with the average values of the diagnostic indicators (Y_{1}’, Y_{2}’, Y_{3}’) for various infections, and conduct a comparison in sign and magnitude, and by the greatest coincidence of the values of the diagnostic indicators of the subject with the average values of the diagnostic indicators for one of urogenital diseases diagnosed it urogenital disease using the following average values of diagnosis:

for chlamydia: Y_{1}’=-2, Y_{2}’=-0,1, Y_{3}’=-0,2;

for Mikael is smasa: Y_{
1}’=2, Y_{2}’=0,8, Y_{3}’=-0,04;

for ureaplasmosis: Y_{1}’=2, Y_{2}’=-1, Y_{3}’=0,02;

healthy: Y_{1}’=-2, Y_{2}’=0,1, Y_{3}’=0,2.

The proposed method was developed by the authors by means of mathematical processing using multifactor analysis of variance using the software “Statistica 1.5” large sample of blood biochemical parameters of healthy volunteers and patients following urogenital infections: chlamydia, mycoplasmosis and a ureaplasmosis.

All were originally studied 34 biochemical indicator of blood, but it was found that the informative value of the diagnosis and differential diagnosis are only four levels in the blood: alpha 1 globulins, beta globulins, gamma globulins and total bilirubin. For each of these four indicators was carried out determination of the coefficients of a multivariate analysis of variance using three well-known models of data by this method using the computer program Statistica 1.5”.

After processing a large set of indicators globulin and total bilirubin in patients with urogenital infections were obtained quantitative values of the coefficients of a multivariate analysis of variance, and because the treatment was carried out on three well-known models multifactor variance is ion analysis, for each model were obtained its value ratios. In the following table 1 shows the values of the coefficients obtained after data processing. Factor values were originally calculated with accuracy to six decimal, but then for convenience of calculations, the values were rounded. Rounded values are shown in table 1 in parentheses.

Table 1 The standard coefficients of the discriminant analysis for the experimental sample | |||

Indicators | The handling model 1 | The handling model 2 | The processing model 3 |

Globulins alpha 1 (B_{1}) | -0,96465 (-1) | 1,010827(1) | -0,099244 (-0,1) |

The beta globulins (_{2}) | -0,585963 (minus 0.6) | -0,168051 (-0,2) | 0,653879 (0,7) |

The gamma globulins (_{3}) | -0,581679 (minus 0.6) | -0,066410 (-0,1) | 0,030187(0,03) |

The total bilirubin (_{4}) | -0,340574 (-0,3) | -0,076411 (-0,1) | -0,895842 (-0,9) |

Then, considering the obtained values of the coefficients of a multivariate analysis of variance tables 1 if computer processing of the results of blood tests large sample of healthy individuals, as well as ill the x chlamydia,
bioplasma and anaplasmosis were obtained average values of diagnosis (Y_{1}’, Y_{2}’, Y_{3}’) for each group surveyed (healthy, chlamydia, mycoplasmosis, ureaplasmosis), which are listed below in table 2, and, also, as in table 1, first, the exact values of parameters, and then, in parentheses, rounded (table 2)

Table 2 Average values of diagnosis (Y _{1}’, Y_{2}’, Y_{3}’for various diseases | |||

The group of patients | Y_{1}’ | Y_{2}’ | Y_{3}’ |

The control group (healthy) | -1,78066 (-2) | 0,11802(0,1) | 0,239081 (0,2) |

Patients chlamydia | -2,21248 (-2) | -0,11696 (-0,1) | -0,215267 (-0,2) |

Patients with Mycoplasma | 2,16336(2) | 0,84794 (0,8) | -0,041503 (-0,04) |

Patients ureaplasmosis | 2,21792(2) | -1,02909 (-1) | 0,021441 (0,02) |

We checked the accuracy of the system and, through an amendment of Bonferroni, narrowed the range of necessary biochemical studies.

Next, using the developed coefficients (table 1) look no further than the biochemical levels in the blood, using the standard discriminant analysis (Statistica 1.5") to conduct a rapid diagnosis and differential diagnosis of the above diseases. For this purpose, the coefficients (table 1) and specific biochemical parameters are entered into the computer, and obtained after processing the values are compared with the average values presented in table 2.

1. For rapid diagnosis and differential diagnosis of diseases chlamydia, mycoplasmosis and a ureaplasmosis, as well as healthy patients is sufficient to use four values of biochemical parameters of blood (shown in table 1).

2. The developed system provides theoretical accuracy of the correct diagnosis is not below 0,913 1%om confidence level. The actual diagnostic accuracy was 0,975.

The proposed method is as follows. In subjects for analysis take blood from a vein. In the blood determine the value of alpha 1 globulin (X_{1}), beta globulin (X_{2}), gamma globulin (X_{3}) and total bilirubin (X_{4}). Then determine the values of the diagnostic indicators (Y_{1}, Y_{2}, Y_{3}for this particular patient using the computer software Statistica 1.5” (USA).

Then the obtained values of diagnostic indicators for konkretnej the patient (Y_{
1}, Y_{2}, Y_{3}) compare with the average values of the diagnostic indicators (Y_{1}’, Y_{2}’, Y_{3}’) for healthy persons and patients with various urogenital infections using the following average values:

for chlamydia: Y_{1}’=-2, Y_{2}’=-0,1, Y_{3}’=-0,2;

for mycoplasmosis: Y_{1}’=2, Y_{2}’=0,8, Y_{3}’=-0,04;

for ureaplasmosis: Y_{1}’=2, Y_{2}’=-1, Y_{3}’=0,02;

healthy: Y_{1}’=-2, Y_{2}’=0,1, Y_{3}’=0,2.

A comparison of Y_{1}, Y_{2}, Y_{3}the average Y_{1}’, Y_{2}’, Y_{3}’ they come in sign and magnitude. In this case, as a rule, than Y_{1}with Y_{1}’ suggest the possibility of two States and then one of them exclude after comparing Y_{2}with Y_{2}’ and Y_{3}with Y_{3}’ (in this order carry out differential diagnostics).

Examples of specific performance are given in the form of extracts from the records.

Example 1.

Patient M., aged 26, extract from map No. 1. Has addressed to the doctor with complaints of pain in the joints. When examining the content of total bilirubin was 8.6 mmol/l, the percentage of globulins alpha 1-2%, beta - 10,9%, gamma - 18,2%.

To determine the values of the diagnostic indicators for this specific patient (i.e. Y_{1/sub>
, Y2, Y3used the computer program "Statistica 1.5";}

this has resulted in the following values:

Y_{1}=-3 (exactly:-3,05447)

Y_{2}=-2 (exactly:-2,12643)

Y_{3}=-0,2 (exactly:-0,22242)

Next conducted a comparison of the values of Y_{1}with Y_{1}’, Y_{2}with Y_{2}’ and Y_{3}with Y_{3}’ using the data from table 2 of this application for each possible state.

By comparing the received data with the average values from table 2 we took the patient to the group of patients with chlamydia. Thus, according to the comparison of Y_{1}with Y_{1}’ this patient can be assigned to control group or chlamydia (match the "-"sign), and according to the comparison of Y_{2}with Y_{2}’ and Y_{3}with Y_{3}’ she can't belong to the control group (has a negative value), and in healthy and Y_{2}’and Y_{3}’ positive (i.e., the discrepancy in sign). Next, the results of our study were confirmed by PCR methods, DFA and IFA.

Example 2.

Patient K., aged 27, extract from map No. 8. I went to the doctor for prophylactic examination. In the study of biochemical parameters of blood obtained the following values: the content of total bilirubin of 20.4 µmol/l, globulin alpha 1-2,7%, beta of 12.9% and a gamma of 17.0%. After processing the data, by definition, Y_{1}, Y_{2}, Y_{3}
the results were as follows:

The results of the analysis | Y_{1} | Y_{2} | Y_{3} |

Patient 2 | 2,19004(2) | -1,03276 (-1) | 0,05153(0,05) |

By comparing the received data with the average values from table 2, we predicted the patient ureaplasmosis. Because according to the comparison of Y_{1}with Y_{1}’ we can exclude belonging to the control group and chlamydia (as in the control group and for chlamydia Y_{1}’ negative), and according to the comparison of Y_{2}with Y_{2}’ and Y_{3}with Y_{3}’ cannot be assigned to mycoplasmosis (as at mycoplasmosis Y_{3}’ negative, and Y_{2}’ positive, i.e. the discrepancy in sign). The results of our study were confirmed by ELISA, MIF and bacteriological sowing.

Example 3.

Patient F., 33 years, an extract from map No. 4. I went to the doctor with complaints of allergic reactions. Along with other analyses were conducted biochemical analysis of blood. The study yielded the following values of biochemical parameters of blood: the content of total bilirubin 20 µmol/l, globulin alpha 1-7,5%, beta of 15.2%, gamma 9,0%. After processing the data with the help of our system b is obtained the following results:

The results of the analysis | Y_{1} | Y_{2} | Y_{3} |

Patient 3 | 2,03285 (2) | 1,68617(1,7) | -0,16208 (-0,2) |

By comparing the received data with the average values from table 2, we diagnosed the patient mycoplasmosis. Because according to the comparison of Y_{1}with Y_{1}’ this patient can be attributed to Mycoplasma or anaplasmosis, and according to the comparison of Y_{2}with Y_{2}’ and Y_{3}with Y_{3}’ we have excluded ureaplasmosis. This result is confirmed by PCR, DFA and bacteriological sowing.

Example 4.

Patient A., 32 years, an extract from map No. 5. When examining the content of total bilirubin was 17.0 mmol/l, the percentage of globulins alpha 1-2,4%, beta of 8.9%, gamma 15.6 per cent. After processing the data with the help of our system the following results were obtained:

The results of the analysis | Y_{1} | Y_{2} | Y_{3} |

Patient 4 | -1,31743 (-1,3) | 0,57285 (0,8) | 1,97344(2) |

By comparing the received data with the average values from table 2, we diagnosed that the patient is 4 refers to a group of PRA is almost healthy persons.
Because according to the comparison of Y_{1}with Y_{1}’ this patient can be assigned to control group or chlamydia, and according to the comparison of Y_{2}with Y_{2}’ and Y_{3}with Y_{3}’ he can't be attributed to chlamydia.

Were examined 40 patients with chlamydia, 40 patients with Mycoplasma, 40 patients ureaplasmosis and 40 healthy persons.

The accuracy of coincidence of diagnosis were: 1 0,975% th confidence level.

Sources of information:

1. Adaskevich VP of sexually transmitted Diseases. - 2nd ed., Corr. and extra - Vitebsk: Publishing house of Vitebsk medical Institute, 1997. - 310 C.

2. Mosquitoes FI, Korovkin B.F., Menshikov V.V. Biochemical studies in the clinic. Elista: APT dzhangar, 1998. - 250 S.

3. Malinina EV Comparative assessment of different methods for diagnosis and therapy of urogenital chlamydia trachomatis in women of reproductive age: author. dis. Kida. the honey. Sciences. - Moscow, 1997. - 22 S.

4. Medical laboratory technology and diagnosis: Reference. Medical laboratory technology./Edited by Professor A.I. Karpischenko. - S.-Petersburg: Intermedica, 1999. - 656 C.

5. Nazarenko GI, Kiskun A.A. Clinical evaluation of laboratory results. - M.: Medicine, 2000. - 544 S.

6. January G.A., I. Ilyin. Mycoplasmal and chlamydial infection of the urogenital tract./Issues of diagnosis and treatment is Iya // Clinical pharmacology and therapeutics. - 1996. No. 4. - P.34 - 39.

The method of rapid diagnosis and differential diagnosis of urogenital infections, including the study of blood, wherein the blood of the subject determine the value of alpha 1 globulin, beta globulin, gamma globulin, and total bilirubin, and then determine the value of the diagnostic indicators of the subject (Y_{1}, Y_{2}, Y_{3}) using the computer program Statistica 1.5”by entering into the computer the values of X_{1}, X_{2}, X_{3}and X_{4}where

X_{1}- the value of the alpha 1 globulin;

X_{2}the value of beta globulin;

X_{3}the value of gamma globulin;

X_{4}- the value of total bilirubin;

obtained values of the diagnostic indicators for this particular patient (Y_{1}, Y_{2}, Y_{3}) compare with the average values of the diagnostic indicators (Y_{1}’, Y_{2}’, Y_{3}’) for various infections, and conduct a comparison in sign and magnitude, and by the greatest coincidence of the values of the diagnostic indicators of the subject with the average values of the diagnostic indicators for one of urogenital diseases diagnosed it urogenital disease using the following average values of diagnosis:

for mantles the oz: Y_{
1}’ = -2, Y_{2}’ = -0,1, Y_{3}’ = -0,2;

for mycoplasmosis: Y_{1}’ = 2, Y_{2}’ = 0,8, Y_{3}’ = -0,04;

for ureaplasmosis: Y_{1}’ = 2, Y_{2}’ = -1, Y_{3}’ = 0,02;

healthy: Y_{1}’ = -2, Y_{2}’ = 0,1, Y_{3}’ = 0,2.

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