1. No category

IJBAF, January, 2013, 1(1): 1-8
ISSN: 2320 – 0774
INCIDENCE OF BACTERIAL AND ABACTERIAL PROSTATITIS AMONG THE
PROSTATITIS PATIENTS IN THI-QAR PROVINCE SOUTH OF IRAQ
ISSA AWATIF H1, AL-SAADI MOHAMMED AK2 AND ATIYALL SAAD A3
1: Department of Biology, College of Sciences, Basra University
2: Department of Microbiology, College of Medicine, Babylon University
3: Department of Microbiology, College of Medicine, ThiQar University
Corresponding Author: E Mail: [email protected]
ABSTRACT
The prostatitis is a worldwide problem affects man at all ages , the incidence of the types of
prostatitis is remain unclear due to the defects in the methods that differentiate between
bacterial and abacterial prostatitis especially culture. Objective of current study was to
determine the incidence of different types of the prostatitis by using culture and PCR based
technique using universal bacterial primer. Post massage urine specimens were collected
from 60 patients with prostatitis and 20 apparently healthy men, the bacterial infections were
diagnosed by culture method and amplification by using B27f-U1492r universal bacterial
primer. Bacterial prostatitis was diagnosed in 11(18.33%) patients by using culture, while,
23(38.33%) patients gave a positive bacterial signals in PCR with high significant differences
(p< 0.001). The remaining patients 37(61.67%) was suffering from abacterial prostatitis.
Therefore, according to the results above abacterial prostatitis were high prevalent among the
patients. But, bacterial prostatitis has high proportion which is larger than those of
international survey which recorded in some studies.PCR technique has a high specificity and
sensitivity in detection of bacteria in prostatic specimens than culture technique.
Keywords: Bacterial and Abacterial Prostatitis, Thi-Qar and Iraq
INTRODUCTION
Prostatitis is an inflammation of prostate
inflammatory
gland
inflammatory prostatitis [1, 2, 3].
classified
into
four
categories
The
bacterial
infectious caused by bacteria, acute bacterial
chronic
and
second
asymptomatic
including: acute bacterial prostatitis, chronic
prostatitis,
first
and
categories
are
prostatitis/chronic pelvic pain syndrome
prostatitis (ABP) is a serious bacterial
which is may be inflammatory or non-
infection of the prostate. Chronic bacterial
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Issa Awatif H et al
Research Article
prostatitis (CBP) is a rare conditions (<5 %)
from prostatitis with age ranged from ( 24-
usually developed as recurrent urinary tract
78 ) years and (20) apparently healthy male
infections originated from recurrent prostate
as controls in Al-Hussein Teaching Hospital
infections [4, 5, 6]. CP/CPPS represent the
in Thi-Qar province through the period from
most prevalent form of prostatitis that
February 2011 to July 2011.
affects men in all ages, data reveals that 210% of adult men suffer from CP and 15%
Culture
The two urine specimens were inoculated on
of men affected at some point of their lives
culture media and incubated aerobically at
[7, 8], the causes of CP/CPPS are remain
37°C for 24h.The bacterial isolation and
unclear. But, it may be associated with
identification was performed according to
psychological factors and dysfunction in
the diagnostic procedures recommended by
immune,
endocrine
[17, 18] by using the colon morphology and
systems and autoimmune responses to
biochemical tests in API 20E system
prostatic
(Biomerux-France).
neurological
antigens
and
represents
important
causes for CP/CPPS [9, 10, 11]. The loss of
symptoms is the most important feature of
Cat.
IV
of
prostatitis
(Asymptomatic
inflammatory prostatitis), inflammation can
be detected in semen, EPS, urine and
Culture technique can be detects only < 10
% of bacterial pathogens that causes
But,
by
using
molecular
techniques such as PCR this percent is rise
greatly to reach to 77% as mentioned by
some researchers [14, 15, 16] by using
universal and
specific
The DNA was extracted from urine samples
according to the manufacturer’s instructions
of
genomic
supplemented
prostate tissue [12, 13].
prostatitis.
Direct PCR-Based Bacterial Detection
DNA Extraction
bacterial DNA
primers. This study aimed to determine the
incidence of the types of prostatitis by
culture and molecular-based methods.
MATERIALS AND METHODS
Samples
DNA
by
purification
the
Kit
manufacturing
company (Sacase-Italy).
PCR Reaction Assays
The extracted DNA were used for PCR
amplification of 16S rRNA genes using the
bacteria
universal
primers
B27F
AGAGTTTGATCCTGGCTCAG-3)
(5and
U1492R (5-GGTTACCTTGTTACGACTT3) [19]. Bacterial 16S rRNA gene sequences
were amplified with prepared master mix
(premix-Bioneer-Korea), the final mixture
contain 5µl premix , 3 µl upstream primer ,
3 µl downstream primer , 5 µl target DNA
The post massage urine samples (VB3) were
and
4
µl nuclease
free
water.
The
collected from (60) male patients suffering
amplification was performed depending on
2
IJBAF, January, 2013, 1(1)
Issa Awatif H et al
Research Article
the following program : 1 cycle of 96°C for
While PCR revealed that 23out of 60
3 min, 56°C for 25 s and 72°C for 15 s and
specimens (38.33%) from the prostatitis
27 cycles of 96°C for 30 s , 56°C for 25 s ,
patients have a positive bacterial signals
and 72°C for 15 s , followed by 72°C for 10
after amplification by PCR and agarose-gel
min. The PCR products were detected by
electrophoresis
agarose gel electrophoresis.
bacterial prostatitis cases as shown in Table
RESULTS
1, the results B27-U1492 primer were
All specimens 20 (100%) of control were
produced 1500 bp PCR products (Figure 1).
give a negative results in culture and PCR
Abacterial prostatitis was shown in the
methods for bacterial detection. Positive
others patients 37 (61.67%) out of 60
culture results were shown in 11 (18.33%)
prostatitis patients with high significant
of patients including (10) isolates of E. coli
differences (p<0.001) as revealed in Table
and (1) isolate of Enterobacteraerogenes.
2.
that
representing
the
Table 1: The Incidence of Bacterial Infections Among the Patients With Prostatitis According to Culture
PCR Methods
Result Methods
Positive
Negative
Total
No.
%
No.
%
No.
%
Culture
11
18.33
49
81.77
60
100
PCR
23
38.33
37
61.67
60
100
Table 2: The Incidence of Bacterial and Abacterial Prostatitis Depending on PCR Techniques
Types of Cases
Prostatitis
Bacterial
No.
23
%
38.3333
Abacterial
No.
37
%
61.6667
Total
No.
60
%
100
Figure 1: Agarose Gel Electrophoresis of PCR of B27-U1492 Universal 16S rRNA Primer Which Showed
a Product of (1500 bp). L: Ladder 1,2, 3, 4,5,6,7: no. of DNA Samples Were Extracted From Post Massage
Urine Samples Obtained From Urine of Prostatitis Patients , lane 5,6,7 was Gave a Positive Bacterial
Signals to the Used Primer
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IJBAF, January, 2013, 1(1)
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Research Article
DISCUSSION
CP/CPPS such as C. trachomatis and genital
Universal PCR primers used as a tool for the
mycoplasmas , these finding lead to
rapid detection of bacteria in normally
developed
sterile clinical samples and it can be using in
bacteria in development of CP/CPPS even
differentiating
viral
though the inflammatory nature of this
infections. This would confirm the necessity
syndrome . Therefore, this finding should be
for antibiotic treatment and would influence
taken in consideration in any attempts to
patient management. Many researchers have
development of an a new classification
used the 16S rRNA gene as target fornon-
system to differentiate between the types of
culture detection and it has been the most
the prostatitis.
widely
PCR technique is the most sensitive and
bacterial
used
targetfor
from
universal
PCR
hypothesis about the role of
amplification of DNAs from a broad range
specific
of organisms [14, 20]. The B27-U1492 give
bacterial infections including prostatitis due
(1500 bp) products and it represents the
to itsability to detects very small number of
most common used universal primer pair
bacteria in the specimens than others
which devised by [21].
methods [25]. Also, PCR is not affected by
The results of current study revealed that the
the presence of some inhibitors in urine and
incidence of bacterial pathogens was high
the inhibition effects of seminal plasma
among the patients suffering from prostatitis
which may suppress the growth of bacteria
which compatible to the results of many
in culture methods, PCR has ability to
researches [4, 14, 15, 22, 23] that shown
detects
high
bacterial prostatitis
chlamydia and mycoplasmas [15, 26, 16, 27,
depending on PCR technique. This result
28, 29]. Also, many patients received
reflects
antibiotics randomly at any time that may
incidence
the
of
role
of
bacteria
in
the
methods
in
unculturable
the
culture
determination
bacteria
results.
such
of
as
developments of the prostatitis either by
affect
Therefore,
direct effects via their toxins and other
molecular method may help in elimination
virulence factors or indirectly by their
the dependence on cultivation methods
ability to induce an immune response and
especially by PCR to detect universal and
inflammation within the prostate tissues
specific 16SrRNA bacterial primers [30].
which lead cell damage and histological
The results of current study was compatible
changes. Moreover , some researchers [14 ,
to some researches [31, 32, 33] which
24] found the bacterial pathogens in the
mentioned that the E. coli is the most
specimens obtained from the patients with
common cause of bacterial prostatitis
4
IJBAF, January, 2013, 1(1)
Issa Awatif H et al
Research Article
because it accompanied with (> 82 %) of
[3] Nickel
JC,
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LM
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Hennenfent M, Research guidelines
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