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Significance of Hematological Biomarkers in Association to Syphilitic Patients

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 Original Article
J Nepal Health Res Counc 2022 Apr-Jun;20(55):426-30

DOI:

Significance of Hematological Biomarkers in

Association to Syphilitic Patients
Arun Bahadur Chand,1 Ajaya Basnet,2 Lok Bahadur Shrestha,3 Bindu Sen,4 Anamika Priyadarshinee,5 Isha
Amatya,6 Bijendra Raj Rajbanshi7
1
Department of Clinical Laboratory, KIST Medical College and Teaching Hospital, Lalitpur, Nepal, 2Department
of Medical Microbiology, Shi-Gan International College of Science and Technology, Kathmandu, Nepal,
3
School of Medical Sciences and The Kirby Institute, University of New South Wales, Sydney, Australia,
4
Department of Dental, KIST Medical College and Teaching Hospital, Lalitpur, Nepal, 5Department of
Pathology, KIST Medical College and Teaching Hospital, Lalitpur, Nepal, 6Research Section, Nepal Health
Research Council, Kathmandu, Nepal, 7Department of Microbiology, KIST Medical College and Teaching
Hospital, Lalitpur, Nepal.

ABSTRACT

Background: Venereal syphilis is a sexually transmitted disease, involving pathological activities mediating tissue
destruction by extensive tissue necrosis. As such, the goal amongst researchers has been set to the identification of
effective laboratory biomarkers that can reflect the broad spectrum of disease and ultimately aid in timely diagnosis and
effective treatment of syphilis. This research aimed to study the applications of hematological biomarkers associated
with syphilitic patients visiting a tertiary care hospital.
Methods: A retrospective cross-sectional study was conducted in the syphilitic patients attending KIST Medical
College and Teaching Hospital, Lalitpur, Nepal. A total of 25 syphilitic patients and 41 non-syphilitic participants were
included. The rapid plasma reagin test and Treponema pallidum hemagglutination assay were used for the screening and
confirmation of syphilis respectively. The hematological investigation was performed using a hematology analyzer.
Statistical Package for Social Science version 17.0 was used for data analysis. A P value <0.05 was considered significant.
Results: Syphilitic patients showed significantly elevated levels of lymphocytes (39.8±11.5) (p=0.025), monocyte
(1.9±0.8) (p=0.002), mean corpuscular volume (MCV) (92.6±12.9) (p=0.005), and mean corpuscular hemoglobin
(MCH) (31.9±4.6) (p=0.008) and lowered levels of red blood cell (RBC) (4.2±0.3) (p=0.005) and platelets
(237.2±628.6) (p=0.048) as compared to the lymphocytes (32.9±11.9), monocyte (0.6±1.2), MCV (83.9±8.8),
MCH (34.3±1.5), RBC (4.6±0.7), and platelets (280.9±113.3) of the non-syphilitic participants.
Conclusions: The results showed that the elevated levels of lymphocyte, monocyte, MCV, and MCH and lowered
levels of RBC and platelets are highly specific hematological biomarkers for the diagnosis of patients with syphilis.
Keywords: Hematological biomarkers; sexually transmitted disease; syphilis

INTRODUCTION through multiple clinical stages over time, eventually

leading to irreversible neurological or cardiovascular
Characterized as a multistage infectious disease with consequences.6 Additionally, infection due to Treponema
a relapsing and remitting course,1 syphilis is usually pallidum, an etiological agent of syphilis, can cross
transmitted through contact with active primary or the placental barrier of a syphilitic mother from the
secondary lesions of a sexual partner.2 In 2015, the World bloodstream and can infect the fetus resulting in
Health Organization (WHO) estimated that there were congenital syphilis,7 which is also a leading cause of
45.4 million syphilitic people in the world,3 with around spontaneous abortion, stillbirth, and perinatal mortality
107,000 deaths.4,5 in many of the countries.8,9

Without treatment, the condition can progress This study aimed to provide a comprehensive overview on

Correspondence: Ajaya Basnet, Department of Medical Microbiology, Shi-Gan

International College of Science and Technology, Kathmandu, Nepal. Email:xlcprk@
gmail.com, Phone: +9779841170737.

426 JNHRC Vol. 20 No. 2 Issue 55 Apr - Jun 2022

 Significance of Hematological Biomarkers in Association to Syphilitic Patients

understanding the significance of altered hematological records were correctly obtained through communication
biomarkers to improve the diagnosis of syphilitic with involved health-care workers or patients and
patients. patient parties. After complete and detailed information
was obtained, it was recorded in a customized data
METHODS collection form. Statistical analysis was performed by
the SPSS program for Windows, version 17.0. Categorical
This was a hospital-based retrospective study conducted variables were presented as absolute numbers (n)
in the Department of Clinical Laboratory, KIST Medical percentage (%), mean ± Standard Deviation (SD). The
College and Teaching Hospital (KISTMCTH), Lalitpur, independent t-test was used for data analysis. A P-value
Nepal from January 2018 to December 2020. The less than 0.05 was considered as a significant difference.
approval was obtained from the Institutional Review
Committee (IRC) of the KISTMCTH (reference number: RESULTS
2077/078/25). Only syphilitic patients who were
tested using rapid plasma regain (RPR) and Treponema Of the total of 66 patients, 37.8% (n = 25) were syphilitic
pallidum hemagglutination assay (TPHA) techniques patients with the mean age ± Standard deviation of
were included, and individuals without syphilis were 29.1±7.7 years and 62.1% (n = 41) were non-syphilitic
excluded. participants with the mean age of 42.9±24.1. While 52%
(n = 13) patients were male from the syphilitic patients,
A total of 66 patients were recruited for the study. There 53.6% (n = 22) patients were male from the non-syphilitic
were 25 syphilitic patients and 41 healthy individuals, participants (Figure 1).
who were considered as the non-syphilitic participants.
Three milliliter whole blood in EDTA was collected for
hematological testing, while 3 milliliter whole blood
in a gel tube was collected for RPR and TPHA assays.
The flocculation card test was used to detect antibodies
produced against antigens released by damaged host cells
in syphilitic patients. In the test, the serum was mixed
with the RPR antigen. If antibodies were present, they
combined with the lipid particles of the antigen, causing
them to agglutinate with showing up as black clumps
against the white card. A rapid diagnostic test kit, TPHA
was used for the diagnosis of syphilis from each serum
sample. In this test, one drop of serum (25 microliters)
followed by one drop of buffer were added to the sample
port. After 15 minutes, the result was detected. Positive
results were indicated by the appearance of two distinct
pinks to deep purple color bands on the device.
Syphilitic
The hematology analyzer (Sysmax XN-550, Japan) was
Non-syphilitic
used for the testing of complete blood cell count (CBC).
The parameters investigated include white blood cell Figure 1. Showing age groups of syphilis subjects and
(WBC), neutrophil, lymphocyte, monocyte, eosinophil, the non-syphilitic participants.
basophil, red blood cell (RBC), hemoglobin, packed cell
volume (PCV), mean corpuscular volume (MCV), mean Higher incidence of syphilis infection (n =17, 68.0%) was
corpuscular hemoglobin (MCH) and mean corpuscular seen in age group of 20-30 years, whereas age groups
hemoglobin concentration (MCHC) and platelet. of 30-40 years and 50-60 years showed an incidence of
28.0% (n = 7) and 4.0% (n = 1) respectively. The youngest
The relevant demographic data and detailed medical syphilitic patient found to be positive in this study was
history of the syphilitic patients were collected from 21 years and the oldest was 56 years (Figure 1).
the hospital record section. Any missing or ambiguous

JNHRC Vol. 20 No. 2 Issue 55 Apr - Jun 2022 427

 Significance of Hematological Biomarkers in Association to Syphilitic Patients

Table1. Hematological biomarkers of the syphilitic Moreover, the results showed a moderate decrease
patients and the non-syphilitic participants. in the MCHC (34.2g/l±2.0) (P>0.05) and a moderate
Group increase in hemoglobin (13.7g/dl±2.5) (P>0.05) of the
syphilis subjects as compared to the MCHC (34.3g/l±1.5)
Parameters Non- P-value
Syphilis syphilitic and hemoglobin (13.7g/dl±2.4) of the non-syphilitic
cases participants. Additionally, there was a significant
WBC (X 109 increase in MCV (92.6%±12.9) and MCH (31.9%±4.6) of
13.3±22.0 9.8±5.6 0.340 the syphilis subjects as compared to MCV (83.9%±8.8)
cells/l)
Neutrophil (%) 57.4±12.0 62.2±13.7 0.158 and MCH (28.9% ±3.5) of the non-syphilitic participants
(Table 1).
Lymphocyte
39.8±11.5 32.9±11.9 0.025
(%)
DISCUSSION
Monocyte (%) 1.9± 0.8 0.6±1.2 0.002
Eosinophil (%) 0.7±2.0 2.4±2.4 0.000 Despite the availability of new diagnostic tests and
Basophil (%) 0.0±0.0 0.03± 0.1 0.439 antibiotic therapy, syphilis has re-emerged in several
RBC (X 1012 developed and developing countries, including Nepal.10
4.2± 0.3 4.6± 0.7 0.005
cells/l) Efforts to eliminate syphilis have met with only modest
Hemoglobin success.11 Alternatively, a quantitative measurement
13.7± 2.5 13.7± 2.4 0.996
(g/dl) of disease severity with the use of clinical biomarkers
PCV (%) 39.6±7.1 38.8±6.7 0.674 reflecting pathological development can be of use to
MCV (fl) 92.6±12.9 83.9±8.8 0.005 clinicians for understanding disease prognosis, outcomes,
MCH (pg) 31.9±4.6 28.9±3.5 0.008 and enabling finesse to decision making.12 Therefore, a
study concerning the laboratory biomarkers of a cohort
MCHC (g/l) 34.2±2.0 34.3±1.5 0.767
of 25 patients with syphilis visiting a tertiary care
Platelets (X hospital was conducted. We studied a clear pattern of 13
237.2±628.6 280.9±113.3 0.048
109 cells/l)
hematological biomarkers. To the best of our knowledge,
WBC = White Blood Cell, RBC = Red Blood Cell, PCV = Packed
Cell Volume, MCV = Mean Corpuscular Volume, MCH = Mean this is the first report concerning the significance of
Corpuscular Hemoglobin, MCHC = Mean Corpuscular Hemoglobin hematological biomarkers and their usefulness in the
Concentration. diagnosis of patients with syphilis from Nepal.

In this study, the majority of syphilitic patients belonged

The results showed a decrease in the neutrophil
to the age group of 20-30 years (68.0%), further preceded
(57.4%±12.0) (P>0.05), eosinophil (0.7%±2.0) (P<0.05),
by the age group of 30-40 years (28.0%) and 50-60 years
and basophil (0.0±0.0) (P>0.05) of the syphilis subjects
(4.0%).The finding was in consensus with the report of
as compared to the neutrophil (62.2%±13.7), eosinophil
the Center for Disease Control and Prevention (CDC),13
(2.4%±2.4), and basophil (0.03%±0.1) of the non-
which mentioned the highest incidence of primary and
syphilitic participants. However, there was a substantial
secondary cases of syphilis among persons aged 25–29
increase (P<0.05) in lymphocytes (39.8%±11.5) (P<0.05),
years followed by 30–34 years. A higher incidence of
monocyte (1.9%±0.8) (P<0.05), and PCV (39.6%±7.1)
syphilis infection in such an age group could be attributed
(P>0.05) of the syphilis subjects as compared to
to not being accustomed to using a condom, not thinking
lymphocytes (32.9%±11.9), monocyte (0.6%±1.2), and
of themselves or a partner as being at high risk for
PCV (38.8%±6.7) of the non-syphilitic participants (Table
syphilis, or not considering oral or anal sex as a way of
1).
contracting or transmitting syphilis.14 In contrast to the
Besides, there was a decrease in the value of study by Obeagu et al.,15 this study was predominated
RBC (4.2X1012 cells/L±0.3) (P<0.05) and platelets by males (52.0%) in the incidence of syphilis infection.
(237.2X109cells/l±628.6) (P<0.05) of the syphilis subjects Similarly, CDC discussed the highest rates of syphilis
as compared to the RBC (4.6X1012cells/L±0.7) and among men (18.7 cases per 100,000 males) as compared
platelets (280.9X109cells/l±113.3) of the non-syphilitic to the rate among women (3.0 cases per 100,000
participants. On the other hand, there was an increase females).13 Higher incidence of syphilis in men could be
in WBC (13.3X109cells/l±22.0) (P>0.05) of the syphilitic attributed to the increased practice of men having sex
subjects as compared to WBC (9.8X109cells/l±5.5) of the with men and/or practice of men going to health care
non-syphilitic participants (Table 1). providers to be tested for HIV.16

428 JNHRC Vol. 20 No. 2 Issue 55 Apr - Jun 2022

 Significance of Hematological Biomarkers in Association to Syphilitic Patients

The mean values for hematological parameters such and the emergence and rapid spread of macrolide-
as neutrophil (57.4%±12.0), hemoglobin (13.7g/ resistant Treponema pallidum are reminders that there
dl±2.5), PCV (39.6%±7.1), MCV (92.6%±12.9), and MCH is no room for complacency. Lack of ability to study
(31.9%±4.6) from this study were in correspondence with other hematological biomarkers such as cytokines,
the study done by Obeagu et al.,15 which reported the immunoglobulin, and involvement of fewer study
mean values of neutrophil, hemoglobin, PCV, MCV, and subjects were the major limitations of the present study.
MCH as 4.5%±0.6, 14.0g/dl±0.5, 42.0%±3.2, 89.9fl±10,
and 29.9pg±5.6 respectively. Similarly, decreased levels CONCLUSIONS
of hemoglobin (1.9g/dL) and neutrophils (23%) were
also reported by a study by Lee et al. Besides, the mean The result showed that syphilis and the ensuing
values for the parameters such as WBC (13.3X109cells/ immune response can have a tremendous impact on
l±22.0), monocyte (1.9%±0.8), eosinophil (0.7%±2.0), the hematopoietic process and the persistence of
and RBC (4.2X1012cells/L±0.3) from the study were such responses may result in chronic and detrimental
in contrast with the result of the research conducted inflammatory feedback to the hematopoietic system.
by Obeagu et al.,15 which reported the mean values The parameters such as lymphocytes, monocytes,
for WBC, monocyte, eosinophil, basophil, and RBC as eosinophil, RBC, MCH, and platelets are significant
4.5cells/l±0.6X109, 30.6%±7.2%, 1.0%±0.1, 0.1%±0.1, hematological biomarkers for syphilis prognosis and
and RBC 4.7cells/l±0.3 X 1012 respectively. The findings diagnosis as compared to other parameters.
of the study coincide with the report of Hira et al.,18
and Kim et al.,18 which mentioned that almost 90% of CONFLICT OF INTEREST
children with early congenital syphilis diagnosed were
None
anemic. Additionally, the findings from our study were
in contrast to the study of Lee et al.,17 who reported the
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