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Virology Section
Laboratory & B.B.
Arar Central Hospital
Northern Borders
Saudi Arabia
Fadi Alanazi
A Laboratory Specialist
BScin BiomedicalScience(UK)
AUDin Laboratory Sciences
• Enveloped, spherical, about 50 nm in diameter .
• The surface proteins are arranged in an icosahedral-like symmetry
• The virus is inactivated by ether, sodium dexoxycholate and chloroform.
VIRION
Family: Flaviridae
Genus: Flavivirus
Species Zika Virus
TAXONOMY
Group IV; ssRNA positive-strand viruses
Nucleotide (top) and amino acid (bottom) , the potential glycosylation sites of the MR 766
(Uganda, 1947) and the IbH 30656 (Nigeria, 1968) strains. Deletions are indicated by
dashes. The ‘‘N’’ at position 467 of the P6-740 strain (Malaysia, 1966) represents an equally
weighted double population of the nucleotides ‘‘C’’ and ‘‘T’’. This translates to an ‘‘X’’ at
position 165 of the amino acid alignment.
(Haddow et al., 2012)
The primer pairs targeting the E gene were
ZIK-ES1 (TGGGGAAAYGGDTGTGGACTYTTTGG)/
ZIK-ER1 (CCYCCRACTGATCCRAARTCCCA) and
ZIK-ES2 (GGGAGYYTGGTGACATGYGCYAAGTT)/
ZIK-ER2 (CCRATGGTGCTRCCACTCCTRTGCCA).
The primer pairs for NS3 amplification were
ZIK-NS3FS (GGRGTCTTCCACACYATGTGGCACGTYACA)/
ZIK-NS3FR (TTCCTGCCTATRCGYCCYCTCCTCTGRGCAGC) and
ZIK-X1 (AGAGTGATAGGACTCTATGG)/
ZIK-X2 (GTTGGCRCCCATCTCTGARATGTCAGT).
RT-PCR with specific primers derived from published ZIKV sequences.
Adenine= thymine, cytosine = guanine
(Grard et al., 2014)
REPLICATION
CYTOPLASMIC nislammam, NUCLEAR nistcesni
1. Attachement (Viral E protein to the host receptors (
2. Fusion of RNA genome into cytoplasm
The +ssRNA is translated into polyproteins
Replication takes place at the surface of endoplasmic reticulum in cytoplasmic
viral factories .
A dsRNA genome is synthesized from the genomic ssRNA .)+(
The dsRNA genome is transcribed/replicated thereby providing
viral mRNAs/new ssRNA)+( genomes.
3. Virus assembly occurs at the endoplasmic reticulum
Then transported to the Golgi apparatus.
4. Release of new virions by exocytosis.
First isolated from Rhesus
monkey (Macaca mulatta)
in Zika forest in Uganda.
Global map of Zika virus infection
Some studies have demonstrated that ZIKV is endemic to Africa and Southeast Asia
(Grard et al., 2014)
Before 2007, few cases of human infection with ZIKV had been reported.
In 2007, an epidemic of ZIKV infection in humans occurred in Yap, Federated States
of Micronesia, in the Pacific region. A seroprevalence survey determined that ≤70%
.of the population had been infected
During 2007–2013, the few cases of infection with ZIKV reported were in travelers
returning from Africa or Southeast Asia
In May 2015, the WHO confirmed the first local transmission of Zika virus in the
Americas in Brazil then spread rapidly to other countries in the Americas.
In January 29, 2016, local transmission has been detected in at least 22 countries or
territories, including the Commonwealth of Puerto Rico and the U.S. Virgin Islands.
Zika virus can infect pregnant women in all three trimesters
Meaney-Delman et al, 2016
Sign and Symptoms
In humans, ZIKV infection is characterized by:
- Mild fever (37.8°C–38.5°C);
- Joint pain( notably of small joints of hands and feet)
- Muscle pain
- Headache
- Conjunctivitis
- Rash
- CBC Test
(mild leukopenia and thrombocytopenia associated with
activated lymphocytes)
ZIKV infection
Believed to be asymptomatic or mildly symptomatic
in most cases. Thus, Zika can be misdiagnosed during
the acute (viremic) phase because of nonspecific influenza like
signs and symptoms. Hemorrhagic signs have not been
reported in ZIKV-infected patients
(Gourinat et al, 2016)
Transient infection in Monkeys, Human, maybe rodents .
Life-long infection in Mosquitoes from the Aedes suneg.
Keratinocytes and dendritic cells at site of inoculation
then spread to lymph nodes and bloodstream.
ASSOCIATED DISEASES:
Adult-> nialliuG-erraBemordnys (Suspected)
Pregnant women- >nrobweNylahpecorcim(detcepsus(
NATURAL HOSTS
TROPISM
- Intellectual disability (decreased ability to learn and function in daily life)
- Problems with movement and balance
- Feeding problems, such as difficulty swallowing
- Hearing loss
- Vision problems
Microcephaly has been linked with the following problems:
Zika is primarily transmitted through the bite of infected Aedes mosquitoes
It can also be transmitted from a pregnant mother to her baby during pregnancy or around
the time of birth
Sexual transmission was reported in two case reports.
There is a potential risk of ZIKV transfusion-derived transmission
TRANSMISSION
There is no evidence that women can transmit Zika virus to their sex partners.
CDC/MMWR Last Update Feb 2016
Guidelines for Breastfeeding for
Mothers with Zika Virus Infection
Zika virus RNA has been identified in breast milk, but attempts to culture the virus have
been unsuccessful
No cases of Zika virus infection associated with breastfeeding have been reported.
CDC encourages mothers with Zika virus infection to breastfeed their infants.
CDC/Morbidity and Mortality Weekly Report (MMWR)Last Update Feb 2016
What is the treatment for Zika?
There is no vaccine or specific medicine to treat Zika virus infections.
Treat the symptoms:
• Get plenty of rest.
• Drink fluids to prevent dehydration.
• Take medicine such as acetaminophen to reduce fever and pain.
• Do not take aspirin or other non-steroidal anti-inflammatory drugs.
NB: If you are taking medicine for another medical condition, talk to your Doctor
before taking additional medication.
What can people do to prevent becoming infected with Zika?
• protect yourself and your family from mosquito bites
• Stay in places with air conditioning or that use window and door screens to
keep mosquitoes outside.
• If you are also using sunscreen, apply sunscreen before applying insect repellent
• If you have a baby or child:
Dress your child in clothing that covers arms and legs, or
Cover crib, pushchair, and baby carrier with mosquito netting.
Are you immune for life once infected?
Once a person has been infected, he or she is likely to be protected from future infections.
Can a previous Zika infection cause a woman who later gets
pregnant to have a baby with microcephaly?
Currently, there is no evidence to suggest that Zika virus, after it is cleared from the blood,
poses a risk of birth defects for future pregnancies.
Zika virus usually remains in the blood of an infected person for about a week.
• Detection of virus RNA in Samples by using reverse transcription PCR (RT-PCR).
• Nucleic Acid Amplification Test.
• Serological tests, including immunofluorescence assays and ELISA may indicate the
presence of anti-Zika virus IgM and IgG antibodies, few laboratories have this ability.
(Gourinat et al, 2016)
ZIKV infection diagnosis relies on serology–which is challenging due to
cross-reactions with other flaviviruses.
Biological confirmation of ZIKV infections
Some facts about sample detection
The suitability of urine samples for diagnosis of ZIKV infection by showing that
ZIKV RNA is detectable in urine at a higher load and with a longer duration than
in serum by real-time RT-PCR.
At the time the rash was observed, viremia was probably decreasing, which
makes detection of virus in serum samples extremely challenging.
Urine samples were positive for ZIKV >10 days after onset of disease, which
was a notably longer period than for serum samples
(Gourinat et al, 2016)
The virus is present in semen longer than in blood. How long? At least 2
weeks after symptoms of infection began.
The use of saliva sample increased the rate of molecular detection of ZIKV at
the acute phase of the disease but did not enlarge the window of detection of
ZIKV RNA. Saliva was of particular interest when blood was difficult to collect
(children and neonates especially).
(Musso et al, 2015)
Some facts about sample detection
• Saliva or urine samples collected during the first 3 to 5 days after
symptom onset.
• Serum collected in the first 1 to 3 days.
Some facts about sample Collection
(WHO/ Western Pacific Region, 2016)
What should be done?
• Alert healthcare providers and the public about Zika.
• Post travel notices and other travel-related guidance.
• Provide regional laboratories with diagnostic tests such as Damma regional lab.
• Detect and report cases, which will help prevent further spread.
References
1. Besnard M, Lastere S, Teissier A, Cao-Lormeau V, Musso D. Evidence of perinatal
transmission of Zika virus, French Polynesia, December 2013 and February 2014. Euro
Surveill 2014;19:20751.
2. Haddow AD, Schuh AJ, Yasuda CY, Kasper MR, Heang V, Rekol Huy, et al. Genetic
characterization of Zika virus strains: geographic expansion of the Asian lineage. PLoS
Negl Trop Dis. 2012;6:e1477. http://dx.doi.org/10.1371/ journal.pntd.0001477
3. Grard G, Caron M, Mombo IM, Nkoghe D, Mboui Ondo S, Jiolle D, et al. (2014) Zika
Virus in Gabon (Central Africa) – 2007: A New Threat from Aedes albopictus? PLoS Negl
Trop Dis 8(2): e2681. doi:10.1371/journal.pntd.0002681
4. Gourinat AC, O’Connor O, Calvez E, Goarant C, Dupont-Rouzeyrol M 2015.
Detection of Zika virus in urine. Emerg Infect Dis 21: 84-86
5. Musso D , Roche C, Robin E, Teissier A, Cao-Lormeau VM (2015). Detection of Zika
virus in saliva. Pub|Med, (68:53-5).
6. ViralZone:www.expasy.org/viralzone, SIB Swiss Institute of Bioinformatics

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Zika virus disease 2016 Updated Version!

  • 1. Virology Section Laboratory & B.B. Arar Central Hospital Northern Borders Saudi Arabia Fadi Alanazi A Laboratory Specialist BScin BiomedicalScience(UK) AUDin Laboratory Sciences
  • 2. • Enveloped, spherical, about 50 nm in diameter . • The surface proteins are arranged in an icosahedral-like symmetry • The virus is inactivated by ether, sodium dexoxycholate and chloroform. VIRION
  • 3. Family: Flaviridae Genus: Flavivirus Species Zika Virus TAXONOMY Group IV; ssRNA positive-strand viruses
  • 4. Nucleotide (top) and amino acid (bottom) , the potential glycosylation sites of the MR 766 (Uganda, 1947) and the IbH 30656 (Nigeria, 1968) strains. Deletions are indicated by dashes. The ‘‘N’’ at position 467 of the P6-740 strain (Malaysia, 1966) represents an equally weighted double population of the nucleotides ‘‘C’’ and ‘‘T’’. This translates to an ‘‘X’’ at position 165 of the amino acid alignment. (Haddow et al., 2012)
  • 5. The primer pairs targeting the E gene were ZIK-ES1 (TGGGGAAAYGGDTGTGGACTYTTTGG)/ ZIK-ER1 (CCYCCRACTGATCCRAARTCCCA) and ZIK-ES2 (GGGAGYYTGGTGACATGYGCYAAGTT)/ ZIK-ER2 (CCRATGGTGCTRCCACTCCTRTGCCA). The primer pairs for NS3 amplification were ZIK-NS3FS (GGRGTCTTCCACACYATGTGGCACGTYACA)/ ZIK-NS3FR (TTCCTGCCTATRCGYCCYCTCCTCTGRGCAGC) and ZIK-X1 (AGAGTGATAGGACTCTATGG)/ ZIK-X2 (GTTGGCRCCCATCTCTGARATGTCAGT). RT-PCR with specific primers derived from published ZIKV sequences. Adenine= thymine, cytosine = guanine (Grard et al., 2014)
  • 6. REPLICATION CYTOPLASMIC nislammam, NUCLEAR nistcesni 1. Attachement (Viral E protein to the host receptors ( 2. Fusion of RNA genome into cytoplasm The +ssRNA is translated into polyproteins Replication takes place at the surface of endoplasmic reticulum in cytoplasmic viral factories . A dsRNA genome is synthesized from the genomic ssRNA .)+( The dsRNA genome is transcribed/replicated thereby providing viral mRNAs/new ssRNA)+( genomes. 3. Virus assembly occurs at the endoplasmic reticulum Then transported to the Golgi apparatus. 4. Release of new virions by exocytosis.
  • 7. First isolated from Rhesus monkey (Macaca mulatta) in Zika forest in Uganda.
  • 8. Global map of Zika virus infection
  • 9. Some studies have demonstrated that ZIKV is endemic to Africa and Southeast Asia (Grard et al., 2014) Before 2007, few cases of human infection with ZIKV had been reported. In 2007, an epidemic of ZIKV infection in humans occurred in Yap, Federated States of Micronesia, in the Pacific region. A seroprevalence survey determined that ≤70% .of the population had been infected During 2007–2013, the few cases of infection with ZIKV reported were in travelers returning from Africa or Southeast Asia
  • 10. In May 2015, the WHO confirmed the first local transmission of Zika virus in the Americas in Brazil then spread rapidly to other countries in the Americas. In January 29, 2016, local transmission has been detected in at least 22 countries or territories, including the Commonwealth of Puerto Rico and the U.S. Virgin Islands. Zika virus can infect pregnant women in all three trimesters Meaney-Delman et al, 2016
  • 11.
  • 12.
  • 13. Sign and Symptoms In humans, ZIKV infection is characterized by: - Mild fever (37.8°C–38.5°C); - Joint pain( notably of small joints of hands and feet) - Muscle pain - Headache - Conjunctivitis - Rash - CBC Test (mild leukopenia and thrombocytopenia associated with activated lymphocytes) ZIKV infection Believed to be asymptomatic or mildly symptomatic in most cases. Thus, Zika can be misdiagnosed during the acute (viremic) phase because of nonspecific influenza like signs and symptoms. Hemorrhagic signs have not been reported in ZIKV-infected patients (Gourinat et al, 2016)
  • 14. Transient infection in Monkeys, Human, maybe rodents . Life-long infection in Mosquitoes from the Aedes suneg. Keratinocytes and dendritic cells at site of inoculation then spread to lymph nodes and bloodstream. ASSOCIATED DISEASES: Adult-> nialliuG-erraBemordnys (Suspected) Pregnant women- >nrobweNylahpecorcim(detcepsus( NATURAL HOSTS TROPISM
  • 15.
  • 16.
  • 17. - Intellectual disability (decreased ability to learn and function in daily life) - Problems with movement and balance - Feeding problems, such as difficulty swallowing - Hearing loss - Vision problems Microcephaly has been linked with the following problems:
  • 18. Zika is primarily transmitted through the bite of infected Aedes mosquitoes It can also be transmitted from a pregnant mother to her baby during pregnancy or around the time of birth Sexual transmission was reported in two case reports. There is a potential risk of ZIKV transfusion-derived transmission TRANSMISSION There is no evidence that women can transmit Zika virus to their sex partners.
  • 20. Guidelines for Breastfeeding for Mothers with Zika Virus Infection Zika virus RNA has been identified in breast milk, but attempts to culture the virus have been unsuccessful No cases of Zika virus infection associated with breastfeeding have been reported. CDC encourages mothers with Zika virus infection to breastfeed their infants. CDC/Morbidity and Mortality Weekly Report (MMWR)Last Update Feb 2016
  • 21. What is the treatment for Zika? There is no vaccine or specific medicine to treat Zika virus infections. Treat the symptoms: • Get plenty of rest. • Drink fluids to prevent dehydration. • Take medicine such as acetaminophen to reduce fever and pain. • Do not take aspirin or other non-steroidal anti-inflammatory drugs. NB: If you are taking medicine for another medical condition, talk to your Doctor before taking additional medication.
  • 22. What can people do to prevent becoming infected with Zika? • protect yourself and your family from mosquito bites • Stay in places with air conditioning or that use window and door screens to keep mosquitoes outside. • If you are also using sunscreen, apply sunscreen before applying insect repellent • If you have a baby or child: Dress your child in clothing that covers arms and legs, or Cover crib, pushchair, and baby carrier with mosquito netting.
  • 23. Are you immune for life once infected? Once a person has been infected, he or she is likely to be protected from future infections.
  • 24. Can a previous Zika infection cause a woman who later gets pregnant to have a baby with microcephaly? Currently, there is no evidence to suggest that Zika virus, after it is cleared from the blood, poses a risk of birth defects for future pregnancies. Zika virus usually remains in the blood of an infected person for about a week.
  • 25. • Detection of virus RNA in Samples by using reverse transcription PCR (RT-PCR). • Nucleic Acid Amplification Test. • Serological tests, including immunofluorescence assays and ELISA may indicate the presence of anti-Zika virus IgM and IgG antibodies, few laboratories have this ability. (Gourinat et al, 2016) ZIKV infection diagnosis relies on serology–which is challenging due to cross-reactions with other flaviviruses. Biological confirmation of ZIKV infections
  • 26. Some facts about sample detection The suitability of urine samples for diagnosis of ZIKV infection by showing that ZIKV RNA is detectable in urine at a higher load and with a longer duration than in serum by real-time RT-PCR. At the time the rash was observed, viremia was probably decreasing, which makes detection of virus in serum samples extremely challenging. Urine samples were positive for ZIKV >10 days after onset of disease, which was a notably longer period than for serum samples (Gourinat et al, 2016)
  • 27. The virus is present in semen longer than in blood. How long? At least 2 weeks after symptoms of infection began. The use of saliva sample increased the rate of molecular detection of ZIKV at the acute phase of the disease but did not enlarge the window of detection of ZIKV RNA. Saliva was of particular interest when blood was difficult to collect (children and neonates especially). (Musso et al, 2015) Some facts about sample detection
  • 28. • Saliva or urine samples collected during the first 3 to 5 days after symptom onset. • Serum collected in the first 1 to 3 days. Some facts about sample Collection (WHO/ Western Pacific Region, 2016)
  • 29. What should be done? • Alert healthcare providers and the public about Zika. • Post travel notices and other travel-related guidance. • Provide regional laboratories with diagnostic tests such as Damma regional lab. • Detect and report cases, which will help prevent further spread.
  • 30. References 1. Besnard M, Lastere S, Teissier A, Cao-Lormeau V, Musso D. Evidence of perinatal transmission of Zika virus, French Polynesia, December 2013 and February 2014. Euro Surveill 2014;19:20751. 2. Haddow AD, Schuh AJ, Yasuda CY, Kasper MR, Heang V, Rekol Huy, et al. Genetic characterization of Zika virus strains: geographic expansion of the Asian lineage. PLoS Negl Trop Dis. 2012;6:e1477. http://dx.doi.org/10.1371/ journal.pntd.0001477 3. Grard G, Caron M, Mombo IM, Nkoghe D, Mboui Ondo S, Jiolle D, et al. (2014) Zika Virus in Gabon (Central Africa) – 2007: A New Threat from Aedes albopictus? PLoS Negl Trop Dis 8(2): e2681. doi:10.1371/journal.pntd.0002681 4. Gourinat AC, O’Connor O, Calvez E, Goarant C, Dupont-Rouzeyrol M 2015. Detection of Zika virus in urine. Emerg Infect Dis 21: 84-86 5. Musso D , Roche C, Robin E, Teissier A, Cao-Lormeau VM (2015). Detection of Zika virus in saliva. Pub|Med, (68:53-5). 6. ViralZone:www.expasy.org/viralzone, SIB Swiss Institute of Bioinformatics

Editor's Notes

  1. The virion RNA is infectious and serves as both the genome and the viral messenger RNA. The whole genome is translated in a polyprotein 3,419 aa long, which is processed co- and post-translationally by host and viral proteases.
  2. Guillain–Barré syndrome (GBS) is a rapid-onset muscle weakness as a result of damage to the peripheral nervous system. Many experience changes in sensation or develop pain, followed by muscle weakness beginning in the feet and hands. The symptoms develop over half a day to two weeks. During the acute phase, the disorder can be life-threatening with about a quarter developing weakness of the breathing muscles and requiring mechanical ventilation.
  3. Amniocentesis is a diagnostic procedure performed by inserting a hollow needle through the abdominal wall into the uterus and withdrawing a small amount of fluid from the sac surrounding the fetus.
  4. Can a previous Zika infection cause a woman who later gets pregnant to have a baby with microcephaly? Currently, there is no evidence to suggest that Zika virus, after it is cleared from the blood, poses a risk of birth defects for future pregnancies. Zika virus usually remains in the blood of an infected person for about a week