Molecular Detection of Molluscum contagiosum virus (MCV) from Patients of Basra Province / Iraq

Molecular detection of
Molluscum contagiosum virus (MCV)
from patients of Basrah province / Iraq
Mohammed Abdullah Gatea
B.S.C Biology ; M.S.C Virology
Assis.Lec. Mohammed A. Gatea

Introduction
Molluscum Contagiosum virus (MCV) is a benign viral infection
that generally affects young children. It is characterized by
smooth, dome-shaped discrete papules (Tom and Friedlander,
2008). The incubation period is variously estimated at 14 days to 6
months (Leslie et al, 2004).
. which comprises 4 genetically subdivided but clinically
indistinguishable MC viral types (Senkevich et al., 1996). MC virus
type I cause the majority (76%-97%) of infection (Scholz et al.,
1989). In patients infected with HIV however, MC virus-2 causes
the majority (60%) of infection (Gold et al., 2006).

Classification
Group : dsDNA viruses
Order : Unassigned
Family : Poxviridae
Subfamily : Chordopoxvirinae
Genus : Molluscipoxvirus
Species : Molluscum contagiosum
With for types MCV1, 2, 3, and 4
(Smithson et al., 2017).

General characterization of (MCV)
Structure and composition :
• Complex symmetry
• Largest viruses from (250-300 nm)
• dsDNA
• Brick shaped
• core ( contains nucleic acid )
• Lateral bodies (unknown function )
• Usually enveloped (glycoprotein + phospholipid)
• *peplomers spike such as Haemaglutinin
and Neuraminidase, binding of virus to host
cell.
• Replicate in cytoplasm*

Replication
Attachment of the viral proteins to host cell.
Penetration by Fusion with the plasma membrane .
Un-coating: Core is completely uncoated , viral genome is free in the
cytoplasm.
Early phase: transcribed in the cytoplasm by viral RNA
polymerase.
Intermediate phase: Intermediate expressed, triggering genomic
DNA replication at 100 minutes post-infection.
Late phase: expressed from 140 min to 48 hours post-infection,
producing all structural proteins.
Assembly of progeny virions starts in cytoplasmic viral factories,
producing an spherical immature particle, virus particle matures
into brick-shaped intracellular mature virion (IMV).
IMV released upon cell lysis, or acquire a second double membrane
from trans-Golgi and bud as external enveloped virion (EEV).

Replication cycle

Epidemiology
Approximately 122 million people were affected worldwide by Molluscum
contagiosum as of 2010 (1.8% of the population).
Genetically there are 4 types of Molluscum. MC virus type I cause the
majority of infection (76%-97%) . However, In patients infected with HIV,
MC virus type 2 causes (60%) from the majority of infection (Gold et al.,
2006).Although current statistics are lacking in the United States, the
overall incidence of Molluscum contagiosum accounts for approximately
1% of all diagnosed dermatologic condition (Kyriakis et al ., 2010).

Transmission route
Transmission
route
Direct infection
Sexual contact
Indirect Infection
Autologous
infection
Infection through
swimming poolsAssis.Lec. Mohammed A. Gatea

102 samples
Molecular study
DNA Extraction
PCR and
Sequencing
Histopathological
study (skin)
Clinical study
Lesion
Color, Shape
and Location, ect…

Clinical results
The present study :-
• That the age of the patients ranged between 1-63 years.
• Our results estimated the size of MCV lesions and it
was between 0.25 to 5 mm.
• The number of lesions ranged from 1-4 lesions on the
same patient.
• The color of infection in most samples (100 patients)
was white, while pink color was in a few samples (2
patients).
• Lesion form in many cases was circular shape (90
lesions) , while the irregular form appeared in a few
cases (12 patients)

characterization of study populations

Distribution age groups and gender

Figure show distribution age groups
0
10
20
30
40
50
60
70
80
90
1-10 11-20 21-30 31-40 41-50 50 <
Age rate (year)
Number
Age (year)Assis.Lec. Mohammed A. Gatea

Figure show the percentage of sex for all age groups .
46
3 7 2 3 2
33
2 2 2 0 0
1-10 11-20 21-30 31-40 41-50 50 <
Sex rate Age group
male female
Number
Age (year)Assis.Lec. Mohammed A. Gatea

Results of Molecular Diagnosis
• The DNA of MCV was extracted by using Tissue
Genomic Viral Nucleic Acid Extraction Kit II
(Geneaid / Taiwan ).
• MCV DNA fragment with the designed primer
set1 yielded band corresponding to their molecular
size of approximately 393 bp and with primer set2
yielded band corresponding to their molecular size
of approximately 575 bp

Set 1 and Set 2 Primers sequence

Figure show Agarose gel electrophoresis of PCR
products L = DNA ladder Lane 1,2,4,5,6,7 = positive to
(393bp).

Figure show Agarose gel electrophoresis of PCR products L =
DNA ladder Lane 1,2,3,4,5,6,7 = positive to (575bp).

Sequencing of PCR products
• MCV isolates were submitted to sequencing , this
isolates were successfully sequenced, virus were
identified to species level, the alignments of the
virus isolates were identified and the molecular
identity of the samples performed by multiple
alignment of each sample sequence with NCBI
database using basic local alignment search tool
(BLAST) software

Table show the Nucleotides Sequencing Data for
Isolates

Histopathological result

Conclusion
• The Molluscum contagiosum is a human disease, predominantly
in males.
• The results of this study showed the distribution of cases
according to age groups. And the age group 1-10 y was more
infected than other age groups.
• distribution of the lesion according to their location revealed
that the head is more sensitive to infection than the other body
parts.
• Molluscum virus infections are lead to in important histological
changes in the skin.
• Molecular detection of molluscum virus is the best way to
detection the virus at level of type.

Recommendation
• Identify the dominant type of MCV in Iraq by studying the
distribution of the virus in different areas of Iraq.
• Study the effects of the immunity to virus infection by MCV to
production of vaccine or drug to this infection.
• The study of MCV infection with other viral diseases such as
HIV or hepatitis to find out overlaps induced virus with patients
immunodeficiency.

Molecular Detection of Molluscum contagiosum virus (MCV) from Patients of Basra Province / Iraq

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