Volume 03 Issue 02-2023
58
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
03
ISSUE
02
P
AGES
:
58-63
SJIF
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5.
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ABSTRACT
The frequency of premature rupture of the membranes before the onset of labor, according to various authors, varies
widely from 1 to 19.8% of cases. pathways, impaired uteroplacental circulation and the development of fetal hypoxia.
KEYWORDS
Pregnant women with premature rupture of membranes, genital tract biocenosis, polymerase chain reaction, preterm
birth, vaginal microbiocenosis.
INTRODUCTION
The microflora of the urogenital tract of women is a
collection of microorganisms that causes resistance to
colonization, pH stability of the vaginal mucosa,
participates in the maintenance of local immunity and,
as a result, ensures the div's general resistance to
infections [1,2,3]. The normal microflora of the vagina
of healthy women of reproductive age includes gram-
positive and gram-negative aerobic, facultative aerobic
and obligate anaerobic microorganisms, 95-98% of all
microorganisms are lactobacilli. Normal microflora also
includes genital mycoplasmas with a titer of less than
104 KOE/ml, fungi of the genus Candida with a titer of
Research Article
SIGNIFICANCE OF THE FEMOFLOR TEST IN ASSESSING THE STATE OF
VAGINAL MICROBIOCENOSIS IN PRETERM VAGINAL DISCHARGE
Submission Date:
February 18, 2023,
Accepted Date:
February 23, 2023,
Published Date:
February 28, 2023
Crossref doi:
https://doi.org/10.37547/ijmscr/Volume03Issue02-12
G.M. Kayumova
Department of obstetrics and gynecology Bukhara state medical institute named after Abu Ali ibn Sina,
Uzbekistan
X.N Hamroyev
Department of obstetrics and gynecology Bukhara state medical institute named after Abu Ali ibn Sina,
Uzbekistan
Journal
Website:
https://theusajournals.
com/index.php/ijmscr
Copyright:
Original
content from this work
may be used under the
terms of the creative
commons
attributes
4.0 licence.
Volume 03 Issue 02-2023
59
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
03
ISSUE
02
P
AGES
:
58-63
SJIF
I
MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
(2023:
6.
184
)
OCLC
–
1121105677
Publisher:
Oscar Publishing Services
Servi
less than 103 KOE/ml [4,5,8]. Bacterial vaginosis occurs
as a result of qualitative and quantitative changes in
the microflora of the urogenital tract. Microorganisms
associated with bacterial vaginosis include Gardnerella,
Mobiluncus, Prevotella, Bacteroides, Fusobacterium,
Peptostreptococcus,
Leptotrichia,
Atopobium
vaginae, and others [6,7,9]. The cause of vaginosis can
be infectious-inflammatory diseases, such changes
often lead to the development of complications in the
organs of the reproductive system. [12,14]. Diseases
caused by conditionally pathogenic microflora in the
mother can lead to spontaneous abortions, premature
discharge of amniotic fluid, intra- and postnatal
infection
of
the
fetus,
postpartum
vaginal
inflammation, and even the development of
premature labor. can come [13,15]. The most common
methods of diagnosing infectious and inflammatory
diseases of the genitourinary tract are microscopic
examination of the condition of the vaginal epithelium
and leukocyte reaction, assessment of the composition
and number of microorganisms by microscopic and
bacteriological examination of smear, qualitative
polymerase chain reaction (PCR). microorganisms can
be present both in pathological conditions (significant
amounts) and in normal conditions (limited amounts).
Therefore,
both
qualitative
and
quantitative
characteristics are necessary to assess the state of
biocenosis [10,11].
The purpose of the study is to determine the
qualitative
and
quantitative
composition
of
microorganisms that make up the microbiocenosis of
the genital tract in pregnant women with premature
discharge using PCR.
RESEARCH MATERIALS AND METHODS
Vaginal swabs of 28 pregnant women who came to the
perinatal center with discharge before the period of 26-
34 weeks and 24-36 years of age were studied. The
control group consisted of 11 pregnant women aged
24-36 with a physiologically developing pregnancy of
20-25 weeks. We used PZR (Femoflor-16) and the DT-96
detection amplification reagent kit manufactured by
OOO "NPO DNK-Technology" (Russia) to study the
biocenosis of the urogenital tract in women. Samples
with sufficient number of cells and sufficient total
bacterial mass from the vagina and partial cervix were
used to obtain appropriate results. Samples were
considered in which the DNA content of human cells
was greater than 104 genome-equivalents (GE) in the
sample and the total bacterial mass value was 106 to
109 GE/sample. After amplification, the total bacterial
mass value of Lactobacillus spp. and each of the
conditional-pathogens was detected automatically.
The state of vaginal microbiocenosis was determined
by their ratio calculated using the software.
Quantitative evaluation of Qin microflora was carried
out both in absolute and relative terms. The absolute
indicator is the amount of DNA of the desired
microorganism in the sample, expressed in GE,
expressed as a decimal logarithm - lg. The relative
quantitative indicator of the microorganism was
calculated as the ratio of the amount of the desired
microorganism to the amount of the total bacterial
mass. It is presented in two formats: the decimal log
difference of the number of the respective group of
microorganisms and the total bacterial mass and as a
percentage of the total bacterial mass.
The state of biocenosis was assessed according to the
criteria developed by G.T. Sukhish et al. The
classification of types of biocenosis includes:
•
absolute normocenosis - a variant of biocenosis, in
which 80-100% of the total bacterial mass of
normoflora is: Ureaplasma spp., Mycoplasma spp.
–
less than 104 ge/ml, and fungi of the genus
Candida spp. - less than 103 g / ml;
Volume 03 Issue 02-2023
60
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
03
ISSUE
02
P
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:
58-63
SJIF
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FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
(2023:
6.
184
)
OCLC
–
1121105677
Publisher:
Oscar Publishing Services
Servi
•
conditional normocenosis - a variant of biocenosis,
in which 80-100% of the total bacterial mass of
normoflora is: the number of Ureaplasma spp and
or Mycoplasma spp.
–
more than 104 ge/ml and
Candida spp. - more than 103 g / ml;
•
moderate (aerobic or anaerobic) imbalance - due
to an increase in the percentage of aerobes or
anaerobes,
the
percentage
of
lactobacilli
decreases by 20-80% compared to the total
bacterial mass;
•
clearly expressed (aerobic, anaerobic or mixed)
imbalance (makes 30-50%) - a variant of biocenosis,
in which the share of aerobes or anaerobes reaches
80-100% of the total bacterial mass, and the share
of lactobacilli decreases to 20% or less. Research
results Processed using Statistica 6.0 software.
Differences were considered significant at p<0.05.
Research results. Comparison of indicators of
biocenosis in the compared groups showed that the
proportion of normocenosis in women with premature
amniotic fluid was 2.5 times lower than in the control
group (p=0.0019). Dysbiosis was detected in 18 out of
28 cases (64.3%), and in the control group in 1 out of 11
women
(9.1%;
p=0.0019).
Average
anaerobic,
pronounced aerobic and mixed dysbiosis was found
only in the main group of women (table 1). Anaerobic
bacteria were detected faster than aerobes, which
corresponds to the results obtained earlier.
Table 1
Determination of the type and level of vaginal dysbiosis in the examined groups, abs (%)
Groups
Normosten
osis
Dysbie
Average
Anaerob
ic
Clearly expressed
Mixed
Anaerob
Аerob
Portion of the water out of order (N
= 28)
10 (35,7)
7 (25,0)
7 (25,0)
2 (7,1)
2 (7,1)
Physiological pregnancy (N = 11)
10 (90,9)
0
7 (25,0)
0
0
0,0019
0,0376
0,1362
0,1869
0,1869
Normocenosis was found in 10 cases of women with
premature amniocentesis. Among them, only 1 patient
was diagnosed with absolute normocytosis, and in the
rest, 8 out of 9 cases had conditional normocytosis
with fungi of the genus Ureaplasma (urealyticum +
parvum) and Candida spp. In 1 case, the titer was more
than 104 and 103, respectively. Anaerobic dysbiosis was
detected on average in 7 (25%) women with premature
amniotic fluid. 24-70% of the total bacterial mass in
cervical smears of patients was Lactobacillus spp.
organizes. Eubacterium spp. It was detected in high
titers in 2 pregnant women.
Volume 03 Issue 02-2023
61
International Journal of Medical Sciences And Clinical Research
(ISSN
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VOLUME
03
ISSUE
02
P
AGES
:
58-63
SJIF
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FACTOR
(2021:
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(2022:
5.
893
)
(2023:
6.
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)
OCLC
–
1121105677
Publisher:
Oscar Publishing Services
Servi
A
combination
of
anaerobic
bacteria
and
microorganisms associated with bacterial vaginosis
was found in 5 cases:
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp.+ Candida fungi;
•
Megasphaera
spp./Veillonella
spp./Dialister
spp.
+Sneathia
spp./Leptotrihia spp./Fusobacterium spp.;
•
Mobiluncus spp./Corynebacterium spp.
+ureaplasma (urealyticum+parvum);
•
Megasphaera
spp./Veillonella
spp./Dialister
spp.+
Ureaplasma
(urealyticum+parvum) + Candida fungi;
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp. + Atopobium
vaginae + Candida fungi.
Of the main group, 9 (32.1%) pregnant women,
including 7 (25.0%) had anaerobic dysbiosis, 2 (7.1%) had
aerobic dysbiosis. In severe dysbiosis, Lactobacillus
spp. amount is from 0 to 18% of the total bacterial mass.
The etiological structure of clearly expressed
anaerobic imbalance consists of the following
combinations of pathogens of anaerobic infections
and microorganisms associated with bacterial
vaginosis:
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp.;
•
Megasphaera
spp./Veillonella
spp./Dialister spp. +Atopobium vaginae +
Ureaplasma (urealyticum + parvum);
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp. + Ureaplasma
(urealyticum + parvum);
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp. + Eubacterium
spp.
+
Sneathia
spp./Leptotrihia
spp./Fusobacterium spp. + Ureaplasma
(urealyticum + parvum);
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp. + Eubacterium
spp. + Mobiluncus spp/|Corynebacterium
spp. + Megasphaera spp./Veillonella
spp./Dialister spp.;
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp. + Eubacterium
spp.;
•
Gardnerella
vaginalis/Prevotella
bivia/Porphyromonas spp. + Sneathia
spp./Leptotrihia spp./ Fusobacterium spp.
Streptococcus spp. observed in high titers. In one of
the patients, along with streptococci, we detected
fungi of the genus Candida in the diagnostic titer. In
both cases Lactobacillus spp. not defined. Moderate
aerobic-anaerobic (mixed) dysbiosis was also detected
in 2 patients. In 1 patient with mixed dysbiosis, the
following
were
detected:
Streptococcus
spp./Gardnerella
vaginalis/Prevolla
bivia/Porphyromonas
spp.,
in
another
case
Megasphaera
spp./Veillonella
spp./Dialister
spp./Ureaplasma (urealyticum+parvum).
Normocenosis was found in 10 out of 11 cases (90.9%)
in physiologically developing women of pregnancy. Of
them, absolute normocenosis was observed in 4
women, conditional normocenosis in 6 women.
Among women with conditional normocenosis,
Ureaplasma (urealyticum + parvum) was detected in 4
cases, fungi belonging to the Candida genus were
detected in 1 patient, and both pathogens were
detected in diagnostic titers in 1 pregnant woman.
Severe anaerobic dysbiosis with a physiologically
developing pregnancy from 11 pregnant women It was
found in 1. The etiological structure is represented by
the
following
pathogens:
Gardnerella
Volume 03 Issue 02-2023
62
International Journal of Medical Sciences And Clinical Research
(ISSN
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2771-2265)
VOLUME
03
ISSUE
02
P
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:
58-63
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FACTOR
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vaginalis/Prevotella bivia/Porp Streptococcus spp.,
Gardnerella hyromonas spp., Eubacterium spp.,
Atopobium
vaginae,
Ureaplasma
(urealyticum+parvum).
CONCLUSION
Thus, as a result of the study, the effectiveness of using
a new modern PCR method for the diagnosis of
bacterial vaginosis was shown. The method allows to
determine the number of lactobacilli and bacteria
associated with bacterial vaginosis, anaerobic and
aerobic flora, including difficult-to-cultivate flora, and
to assess their ratio and the state of the biocenosis of
the female genital tract in a short period of time. For
the first time, using the Femoflor test, the microbial
landscape of the genital tract of women with
premature ejaculation was compared with the
microbiocenosis of the genital tract of women with
physiological
pregnancy.
In
64.3%
of
cases
Lactobacillus spp. a decrease in the percentage,
bacterial imbalance was detected in pregnant women
with premature amniocentesis. At the same time, in
women with physiological pregnancy, the decrease of
lactobacilli was detected only by 9.1% (p=0.002), in
most cases, normocytosis was detected. Anaerobic
microorganisms (14 out of 28 or 50%) played the main
role in the structure of dysbiosis in pregnant women
with premature amniocentesis: Gardnerella vaginalis
(17.8%), Candida spp. genus fungi (10.7%), Ureaplasma
(urealyticum + parvum) - 17.8%, Atopobium vaginae
(7.1%). Aerobic dysbiosis was noted only in 2 women
(7.1%), Streptococcus spp. and Candida spp. fungi of
the genus, mixed - observed in 2 (7.1%) cases. The
identified facts confirm the point of view about the
important role of bacterial vaginosis in the genesis of
premature birth, emphasize the need to diagnose
dysbiosis in the restoration of normal microflora.
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