Differential diagnostic technique for generalised form of intrauterine mono- and mixed cytomegalovirus infection in newborns

FIELD: medicine.

SUBSTANCE: seromucoid concentration is measured in supernatant of a biological fluid aspirated from the nasopharynx of the newborns suffering a generalised form of the intrauterine mono-cytomegalovirus infection or mixed cytomegalovirus infection. If the seromucoid concentration is 0.110-0.140 absorbance units, the early stage of the generalised form of the intrauterine mono-cytomegalovirus infection is diagnosed. If the seromucoid concentration is 0.141-0.171 absorbance units, the early stage of the generalised form of the intrauterine mixed cytomegalovirus infection caused by a combination of the cytomegalovirus and type 1 herpes simplex virus is diagnosed.

EFFECT: using the declared method enables the effective differential diagnosis of the generalised form of the intrauterine mono or mixed cytomegalovirus infection in the newborns.

1 tbl, 3 ex

 

The invention relates to medicine, namely to Perinatology and neonatology.

In children of early neonatal age are often diagnosed generalized form of intrauterine mono - and mixed-cytomegalovirus infection accompanied by a systemic inflammatory reaction[1, 2, 4, 7]. With the direct effects of viruses on cells and tissues in serum and other biological fluids are defined markers of inflammation, one of which is seromucoid - glycoprotein related to biokompozitnyh produce proteins [3]. Known method for the diagnosis of generalized forms of intrauterine infection, based on the determination in the amniotic fluid in 10 fields of view of the absolute content of macrophages (M), lymphocytes (L), segmented neutrophil leukocytes (N) and epithelial cells of the fetal skin (e) and calculating the equation - 1,007×M-0,723×N-0,826×L+0,005×e, and if the received rate is below the threshold, equal - 12,51, diagnose generalized form of infection of the fetus [5].

The disadvantage of this method [5] is that it does not allow for differential diagnosis in newborns generalized form of intrauterine mono - and mixed - cytomegalovirus infection.

The proposed method is to ensure the differential diagnosis of generalized FD�we intrauterine mono - and mixed-cytomegalovirus infection in newborns.

The method includes the following techniques:

1. To sterile syringe attached soft catheter [6].

2. A soft catheter is carefully inserted into the nasal cavity and nasopharyngeal space newborn immediately after birth.

3. Syringe slowly nasasyvaût 2.5 to 3 ml of the biological fluid contained in the nasal passages and nasopharynx of a newborn.

4. Obtained 2.5-3 ml of biological fluid nasopharyngeal aspirate was centrifuged in a centrifuge at 1500 rpm for 10 minutes to obtain supernatant her part.

5. To determine seromucoid is taken 0.2 ml of biological fluid aspirate and added 0.2 ml of 0.95% aqueous solution of sodium chloride and 0.2 ml of 0.45 molar sulfosalicylic acid. Within 30 minutes, the mixture was centrifuged at 1500 rpm Then 0.3 ml of the supernatant is added 0.6 ml of 0.85% aqueous solution of sodium chloride and 4 ml of a 0.1% aqueous solution of tannin, mixed and boiled for 30 minutes in an ice bath and poured in a 10 mm cuvette of fotoelektrokalorimetry (FEC). Determined by the rate of extinction at 590 nm (red filter). The standard of comparison is distilled water [3].

6. Intrauterine cytomegalovirus infection is diagnosed by the detection of the titer of IgM antibodies to cytomegalovirus (CMV) 1:200 in umbilical cord blood in newborns, a fourfold growth t�spectra of IgG antibodies to CMV in neonates compared with that of their mothers, for example, 1:200-1:800 using the test systems of the company "VECTOR-BEST" (Russia), and polymerase chain reaction (PCR) markers deoxyribonucleic acid (DNA)-CMV in umbilical cord blood.

7. Intrauterine herpes infection is diagnosed by the detection of the titer of IgM antibodies to herpes simplex virus type 1 (HSV-1 type) 1:200 in umbilical cord blood in newborns, a fourfold rising titer of IgG antibodies to HSV-type 1 in newborns compared with those from their mothers, for example, 1:200-1:800 [2], and also PCR-DNA markers-the herpes simplex virus in cord blood.

8. Conduct differential diagnosis of generalized forms of intrauterine mono - and mixed-cytomegalovirus infection in newborns: when the concentration of seromucoid equal 0,110-0,140 units of optical density (units wholesale. the raft.), diagnose generalized form of intrauterine mono-cytomegalovirus infection; when the concentration of seromucoid equal 0,141-0,171 units wholesale. raft., diagnose generalized form of intrauterine mixed cytomegalovirus infection, caused by a combination of CMV and HSV-1 type.

For the purpose of establishing the significance of the study the concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate for the differential diagnosis of generalized forms of intrauterine mono - and mixed-cytomegalo�rosnay infection, we surveyed 51 newborn. The control group consisted of 30 healthy newborns from mothers with physiological pregnancy. In the first experimental group consisted of 25 newborns with generalized form of intrauterine mono-cytomegalovirus infection. The second experimental group consisted of 26 newborns with generalized form of intrauterine mixed cytomegalovirus infection, caused by a combination of CMV and HSV-1 type. In newborns the first and second experimental groups generalized form of intrauterine mono - and mixed-cytomegalovirus infection diagnosed for 6 hours before the development of clinical disease.

The concentrations of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate in newborns of the control group, the first and second experimental groups are presented in the table.

The table shows that in infants with generalized form of intrauterine mono, cytomegalovirus infection, compared with newborns of the control group, increases the concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate to 0.125±0,001 (0,110-0,140) % opt. raft. (p<0,001).

Comparison of concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate in newborns the first and second experimental groups showed statistically significant �Velicina in newborns with generalized form of intrauterine mixed cytomegalovirus infection due to a combination of CMV and HSV-1 type, to 0,156±0,001 (0,141-0,171) % opt. raft. (p<0,001).

Given the statistical significance of the concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal

Table
The concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate in newborns of control and test groups
Statistical indicatorsThe concentration of seromucoid (unit opt. the raft.)
The control group (n=30)The first experimental group (n=25)The second experimental group (n=26)
M0,0950,1250,156
m0,0030,0030,003
σ0,010,010,01
M±1,5 δ0,080-0,1090,110-0,140 0,141-0,171
p-<0,001<0,001
p1-<0,001
Note: p - significance of differences between parameters in the control, the first and second experimental groups; p1- the level of significance of differences between parameters of the first and second experimental groups.

aspirate in newborns with generalized intrauterine form mono - and mixed-cytomegalovirus infection, we propose to use these indicators as criteria for differential diagnosis of generalized forms of intrauterine mono - and mixed-cytomegalovirus infection (table).

When the concentration of seromucoid in the supernatant portion of the biological fluid aspirate 0,110-0,140% opt. raft. diagnose generalized form of intrauterine mono, cytomegalovirus infection, and when the concentration of seromucoid 0,141-0,171 units wholesale. raft. diagnose generalized form of intrauterine mixed cytomegalovirus infection, caused by a combination of CMV and HSV-1 type.

To illustrate the effectiveness of the proposed method of differential diagnosis of generalized forms of intrauterine mono - and m�FTC cytomegalovirus infection, the following clinical examples.

Example 1

Baby S. was Born from the second pregnancy. The mother was regularly observed in the antenatal clinic, where he was registered with 6 weeks. During the first, second and third trimesters of pregnancy without features. Serologically in the serum of the mother was not determined markers of chronic cytomegalovirus infection, confirmed by the absence of IgG antibodies to CMV. Detected IgG antibodies to HSV-type 1 titer of 1:200. First birth, at term, vaginally. Amniotic fluid light.

Clinical diagnosis mother's first Birth, in time. Amniotomy. Episiotomy.

A girl was born with a mass 3890 g, length 55 cm, head circumference 35 cm and chest 33 cm In the delivery room the Apgar score 8/10 score. In a serological analysis in serum of umbilical cord blood of a newborn was absent IgG antibodies to CMV and IgG antibodies to HSV-type 1 and also not detected markers DNA CMV and HSV DNA.

In umbilical cord blood of a newborn leukocyte count was equal to 10.2x109/L. In the leucocyte count eosinophils was 1%, stab neutrophils - 2%, segmented neutrophils - 57%, lymphocytes - 35% and monocytes - 6%. The concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate obtained immediately after birth, was 0,085% opt. the raft.

Prirozhdennye the child's condition was satisfactory. The skin of the face and torso was pink. The Fontanelle was 1.5×1.5 cm, and was not stressed. The cry of the child was loud, and muscle tone and tendon reflexes are normal. Physiological reflexes were clearly visible. Heart sounds clear, rhythmic to 140 beats per 1 minute. Respiratory rate of 44 to 1 minute. The breath in the lungs puerile, was carried out across all fields. The external genitals are developed correctly on the female type.

The early neonatal period was uneventful. The umbilical cord fell off at 3 days. The child was discharged from the Department in satisfactory condition.

Clinical diagnosis: the neonatal Period.

Example 2

Newborn Y. was Born from the first pregnancy. In female consultations were observed regularly from 6 weeks of gestation. In the first 7 weeks of gestation, the mother showed signs of early preeclampsia (nausea, vomiting up to 3 times a day). In 18 weeks serologically diagnosed exacerbation of chronic cytomegalovirus infection. In a serological analysis of the mother was determined IgM antibodies to CMV 1:200-1:400, and the increase of IgG antibodies to CMV 1:200-1:800 in paired sera. With 25 weeks recorded the symptoms of threatened miscarriage and chronic placental insufficiency, for which she was treated in the hospital. First birth, at term, vaginally. Amniotic fluid light.

p> Clinical diagnosis of the mother: a Birth in the first period. Reactivation of chronic cytomegalovirus infection. Chronic placental insufficiency. Chronic intrauterine fetal hypoxia. The episiotomy. Apiterapia.

A girl was born weighing 3360 g, length 53 cm, head circumference - 35 cm chest 34 see In the delivery room assessment by Apgar 5/7 points. Bacteriological examination of the nasal mucosa microflora was not detected. Serology serum of blood from a vein of the umbilical cord of a newborn found intrauterine cytomegalovirus infection (IgM antibodies to CMV 1:100 and IgG to CMV 1:200). PCR markers of CMV DNA - positive. At birth she the total number of leukocytes was 8.5×109/L. In the leucocyte count of eosinophils was 5%, stab neutrophils - 1%, segmented neutrophils - 40%, lymphocytes 43% and monocytes - 11%. The concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate was 0,132 units wholesale. the raft.

The condition of the newborn closer to satisfactory. Was observed cyanosis of nasolabial triangle, as well as decreased activity and muscle tone. The tendon reflexes of the child have been weakened. Physiological reflexes indistinct. Cardiac sounds are muffled, rhythmic to 140 beats per 1 minute. Respiratory rate 47 in 1 minute. In the lungs revealed�ü the limited areas of weakened breathing. After 6 hours, the condition of the newborn heavy. He also started moaning breathing, acrocyanosis. Heart sounds muffled to 152 in 1 minute. It was noted the difficulty breathing through the nose and wings of the nose. Registered retraction of the intercostal spaces on inspiration. Respiratory rate increased to 62 in 1 minute. Auscultation the breath in the lungs with a hard tone. There wheezing. When disturbed there was an increase of tension of the anterior Fontanelle and its pulsation, as well as a significant decrease in motor activity. Identified electoral alternation of tone in the flexor muscles, turning into cramps. Was observed a pronounced symptom Graefe, as well as nystagmus and strabismus. Notice the asymmetry of muscle tone. Doppler study of cerebral vessels for 3 days showed increased resistance index in the middle cerebral artery to 0.77.

The newborn diagnosis: generalized Intrauterine cytomegalovirus infection. Intrauterine pneumonia. Cerebral ischemia of moderate severity. Hypertension-hydrocephalic syndrome.

Example 3

Newborn K. was Born from the first pregnancy. The mother was observed in the antenatal clinic regularly since 5 weeks. To 8 weeks the pregnancy was showing symptoms of mild preeclampsia (nausea, vomiting up to 1 times sutki). At 22 weeks marked reactivation of chronic cytomegalovirus and herpes infection (HSV-1). Serologically the mother was determined IgM antibodies to CMV 1:400 and the increase of IgG antibodies to CMV 1:200-1:800. Simultaneously detected IgM antibodies to HSV-1 type 1:200 and IgG antibodies to HSV-1 type 1:200-800 in paired sera. At 24 weeks was diagnosed with chronic placental insufficiency, for which he was treated in hospital. First birth at term, vaginally. The amniotic fluid is green.

Clinical diagnosis mother's first Birth, in time. Reactivation of chronic cytomegalovirus-herpes infection. Chronic placental insufficiency. Chronic intrauterine fetal hypoxia. The episiotomy. Apiterapia.

The boy was born with a mass 3230 g, length 52 cm, head circumference - 34 cm chest 34 see In the delivery room the Apgar score 4/7 points. Blood group A(II), RH positive. A newborn's condition at birth moderate severity. The skin of the face, extremities and trunk clean. It was noted cyanosis of nasolabial triangle. Muscle tone is reduced, the tendon reflexes are called weakly. Physiological reflexes are not clearly expressed. Cardiac sounds are muffled, rhythmic to 147 beats per 1 minute. Respiratory rate 45 in 1 minute. The breath in his lungs weakened.

In a serological study saw�rocky of blood from the umbilical vein in the newborn were detectable markers of fetal cytomegalovirus-herpes infection (IgM antibodies to CMV 1:100 and IgG to CMV 1:200 and IgM antibodies to HSV-1 type 1:100 and IgG antibodies to HSV-1 type 1:200). PCR markers for CMV and HSV - positive. The number of leukocytes was $ 9.0×109/L. In the leucocyte count banded neutrophils was - 2%, segmented neutrophils 48%, lymphocytes - 39%, monocytes - 11%. The concentration of seromucoid in the supernatant portion of the biological fluid nasopharyngeal aspirate obtained immediately after birth, was 0,167 units wholesale. the raft.

When examining a child after 6 hours negative dynamics was observed. A newborn's condition with heavy signs of acrocyanosis and frequent regurgitation of aspirated amniotic fluid with meconium. Appeared the asymmetry of the corners of the mouth. On inspection reacted painful scream. Showed a significant decrease in activity, muscle tone, depression of tendon and physiological reflexes. Heart sounds were muffled, rhythmic to 159 beats per 1 minute, and respiratory rate was increased to 61 in 1 minute. Breath was held over much of the surface of the lungs was attenuated with a single wheezing. X-ray examination diagnosed intrauterine pneumonia. Often observed moderately expressed flatulence. Palpation was determined by increasing the tension of the anterior Fontanelle and its ripple. Significantly decreased motor activity. Pay attention ceredo�of tone in the flexor muscles, often turning into convulsions. Pronounced symptom Graefe, and horizontal nystagmus. Was observed asymmetry of muscle tone. Doppler study of cerebral vessels for 3 days showed increased resistance index in the middle cerebral artery to 0.79.

Clinical diagnosis of the newborn: Intrauterine generalized cytomegalovirus-herpes infection (HSV-1). Intrauterine pneumonia. Cerebral ischemia of moderate severity. Hypertension-hydrocephalic syndrome.

The technical result of the invention is as follows.

1. Our proposed method of differential diagnosis of generalized intrauterine mono - and mixed-cytomegalovirus infection in newborns is based on the definition in the supernatant portion of the biological fluid nasopharyngeal aspirate seromukoida: when the concentration of seromucoid equal 0,110-0,140% opt. raft., diagnose generalized form of intrauterine mono, cytomegalovirus infection, and when the concentration of seromucoid equal 0,141-0,171 units wholesale. raft., diagnose generalized form of intrauterine mixed cytomegalovirus infection, caused by a combination of CMV and HSV-1 type.

The method was tested in the obstetric patient Department of the Amur regional clinical hospital during the WA�the study 51 of the newborn and allows in 94.1% of cases differential diagnosis of generalized intrauterine mono - and mixed-cytomegalovirus infection 6 hours before the development of clinical infectious disease.

Used information sources

1. Likhachev A. A., Malakhov, Y. E., Mordovin B. V. To the question of the clinical-morphological forms of generalized cytomegaly//Issues of the PU. Mat. and det. - 1972. - Vol. 17, No. 8. - P. 39-45.

2. Samokhin P. A. Cytomegalovirus infection in children (clinical and morphological aspects). - M.: Medicine, 1987. - 160 p.

3. Samsonov, V. P., Lutsenko M. T., Novik E. V. Diagnostics of various degrees of endotoxemia abscesses in the lungs: methodical recommendations of Ministry of health of Russia, Institute of physiology and pathology of respiration SB AMS USSR. - Blagoveshchensk, 1988. 10 s.

4. Serov V. N., Musykantova B. C., Kalashnikov, V. G. Congenital cyto-megalia - clinical-anatomical shape // Akush. and Hynek. - 1992. - No. 3-7. - S. 33-36.

5. Method for the diagnosis of intrauterine infection of the fetus generalized forms: the patent of the Russian Federation 2205406, IPC G01N 33/487 / N. M. Khamula, T. S. Bystritskaya, Tseluyko S. S.; applicant and patentee of the Amur state medical Academy. - No. 2001118226/14; various types. 02.07.2001; publ. 27.05.2003.

6. Device for aspirating fluids from the nasal cavity and nasopharyngeal space in infants: a patent of the Russian Federation 2392974, IPC A61M 1/00 / I. N. Gorik, M. T. Lutsenko, V. P. Samsonov, L. G. Nachamkin, E. V. Ushakov, N. About. Kostromina; applicant and patentee GU, Dagestan scientific center SB RAMS. - No. 2008152941/14; various types. 31.12.2008; publ. 27.06.2010, bull. No. 18.

7. Fedorova M. V., Serov VN., Strizhakov A. N., Timofeev, T. G. Intrauterine infection // Vestnik Ross. Association of obstetricians and gynecologists. - 1997. - No. 2. Pp. 89-99.

A generalized method of differential diagnosis of intrauterine form mono - and mixed-cytomegalovirus infection in infants with generalized intrauterine form mono - and mixed-cytomegalovirus infection by determining their seromucoid in the supernatant portion of the biological fluid, nasopharyngeal aspirate, characterized in that when the concentration of seromucoid 0,110-0,140 units of optical density diagnose the early stage of the development of generalized forms of intrauterine mono, cytomegalovirus infection, when the concentration of seromucoid 0,141-0,171 units of optical density are diagnosed with early stage generalized form of intrauterine mixed cytomegalovirus infection, due to a combination of cytomegaly virus and herpes simplex type 1.



 

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14 cl, 3 tbl, 4 dwg

FIELD: medicine.

SUBSTANCE: invention represents a method for the prediction of preeclampsia in the second trimester of pregnancy by blood examination, differing by the fact that the activity of acid and neutral proteinases is measured in blood serum of the women 7-8 weeks pregnant; if the activity of acid proteinases is more than 5.6 mcmole/l, while the activity of neutral proteinases is more than 3.9 mcmole/l, the preeclampsia progression in the second trimester of pregnancy is predicted.

EFFECT: higher accuracy and specificity of the method for the prediction of gestational toxicosis.

2 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to a method of automated morphometric myelofibrosis diagnostics. The method essence consists in the fact that overview images of zones with different optical properties with determinable fibrous and heamopoietic properties of a biological tissue are performed. Ratios of areas of the said zones of at least three paraffin cuts of trepanobiopsy samples are calculated. The coefficient (Cop) is calculated as the ratio of the fibrous tissue area to the area of the heamopoietic tissue by formula. If the value Cop ≥14.5%, myelofibrosis is diagnosed.

EFFECT: application of the claimed method makes it possible to increase the accuracy and improve the efficiency of myelofibrosis diagnostics.

7 cl, 1 tbl, 4 dwg, 1 ex

FIELD: medicine, psychiatry.

SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.

EFFECT: more objective prediction of disease development.

3 ex

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