Instituto Valenciano de Microbiología
(IVAMI)

Masía El Romeral
Ctra. de Bétera a San Antonio Km. 0.3
46117 Bétera (Valencia)
Phone. 96 169 17 02
Fax 96 169 16 37
Email: 
www.ivami.com
CIF B-96337217

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Nipah virus - Molecular diagnosis (PCR).
 
Information 16/10/14.

The Nipah virus is named after the place where people first infected (Kampung Sungai Nipah - Nipah River Village, Town Nipah River) were found.
         
The Nipah virus is an emerging virus, first detected between September 1998 and June 1999, in an outbreak that affected people who had a fever of acute onset with encephalitis in Malaysia and Singapore, which affected 283 people, with a fatality rate of 40%. Subsequently, seven outbreaks have been detected in Bangladesh being 2001 and 2007, and several outbreaks in India, some close to the Bangladesh border areas.
It is now recognized that the reservoir of the virus is in fructívoros bats of the genus Pteropus (Pteropus giganteus, a frequent flying fox in Southeast Asia) that have been found infected in Malaysia, Bangladesh and Cambodia. At the outbreak of Malaysia, human outbreak occurred in infected pigs from (most cases occurred in pig keepers), and Singapore outbreak occurred in pig slaughterhouses from Malaysia. Therefore, initially it thought that the reservoir would be in these animals. Outbreaks in Bangladesh has not found an intermediate animal host from which people became infected, and it is admitted that people were infected directly or indirectly from infected bats or other infected people initially. This human transmission has been confirmed outbreaks in Bangladesh.
       
The virus affects the nervous system, lung and kidney. Therefore, most patients develop severe acute encephalitic syndrome and many patients develop respiratory symptoms. Signs and symptoms observed in these patients in order of frequency are: fever (100%), altered mental status (90%), headache (73%), weakness (67%), respiratory distress (69%), cough ( 62%), diarrhea (29%), seizures (23%). You should
be ruled out infection in people from the regions of Southeast Asia where people have found these outbreaks, which have a febrile syndrome with altered state of sensory (awareness - mental), and sometimes with shortness of breath and headaches.
       
In the diagnosis of this infection is necessary
to rule out other infections that can be detected in patients from countries found infection with this virus, such as malaria, Japanese encephalitis, disease West Nile virus (West Nile), Dengue infection Hantavirus, or Leptospira infection.

Microbiological diagnosis - virological of Nipah virus

  • Culture isolation: this virus can be isolated in culture in the laboratory using Vero E6 cells. The virus induces syncytium formation in cell cultures inoculated with samples containing Nipah virus, and can be confirmed by diagnostic methods developed RT-PCR and sequencing. This method is not currently the first diagnostic alternative because primary isolates may require two blind passages to 5 days in culture, before giving such negative virus culture.

  • Molecular diagnosis by RT-PCR with confirmation by sequencing. This method is most recommended for its speed today.

  • Serological diagnosis: detecting IgM or IgG antibodies, only performed on centers for the disease, so it is not a recommended standard method, and at the same time is later than the molecular methods.

Tests in IVAMI

  • Molecular diagnosis.  Recommended method.
  • Culture isolation. Second diagnostic alternative but take longer.

Recommended samples for microbiological diagnosis - virological

  • Central nervous system: cerebrospinal fluid sample, 2 mL in sterile polypropylene vial.
  • Respiratory infection: aspirate sample tube or deposited in sterile polypropylene container.
  • Renal disease: urine sample, 10 mL in container or sterile polypropylene tube.

Preservation and shipment of sample:

  • Refrigerated (preferred) for less than 2 days.
  • Frozen: over 2 days.

Delivery of results

  • Microbiological molecular diagnostics - 48 hours
  • Culture isolation - 5 days.
    
Cost of the test

  • Molecular diagnosis - Seek ivami@ivami.com.
  • Diagnosis by culture (culture isolation) - Seek ivami@ivami.com.