The Laboratorian - Volume 5, Issue 3


The Laboratorian - Volume 5, Issue 3
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January 2014 - Volume 5, Issue 3

Article Index

- Viral Isolation
- Special NBS Tour
- 10 Year Celebration


Influenza testing has been a recurring topic in The Laboratorian since this newsletter launched just after the 2009 H1N1 outbreak. The Viral Isolation team was gracious enough to welcome our virtual tour at the height of their busy season. Read the lead article, "Lab Tour: Viral Isolation," to see how viruses are quickly identified – a crucial step in containing outbreaks. 

Once again, the Texas Newborn Screening program had the privilege of hosting visitors. This time our distinguished guests came from India to learn how our NBS processes can help their fledgling newborn screening program. For details read "Newborn Screening Tour Aids India." 

November 2013 marked 10 years in our new laboratory building. As the saying goes, 'Time flies when you are having fun,' and for long-term laboratorians it does feel like the move wasn’t that long ago. Moving into this building was a staged process, so memories of exact dates are sometimes a bit vague, but I have done my best to re-construct the sequence of events in the article, “Celebrating 10 Years in the Laboratory Building.”

by Jimi Ripley-Black

Lab Tour: Viral Isolation

A laboratorian performs manual RNA Extraction. Photo courtesy of Andrew Vinyard.

The middle of the flu season seems an appropriate time to visit Viral Isolation (VI) to learn about the viruses – including Influenza – that they identify and isolate. VI also tests for Measles, Mumps, Rubella, Varicella-Zoster Virus, Parainfluenza, Respiratory Syncytial Virus, Herpes Simplex Virus 1/2, Adenovirus, Enterovirus and Cytomegalovirus.

Specimens submitted to VI come from doctor’s offices, hospitals, clinics, commercial laboratories, and local health departments from around Texas. As you might imagine, with VI’s involvement in detecting outbreaks, work volume can vary greatly from year to year. The majority of specimens are Influenza Surveillance specimens. Team Lead Crystal Van Cleave listed specimen types and tests with practiced ease.

  • “We receive specimens for Mumps and Middle East Respiratory Syndrome (MERS) Coronavirus for Real-Time RT-PCR testing;
  • General Virus Isolation specimens (submitter thinks that a virus is the cause of illness and may or may not suspect a particular virus, cell culture is performed on these);
  • Reference Culture specimens (submitter has previously performed cell culture and sends us a cell culture isolate for confirmation);
  • And Electron Microscopy.”

All of these tests are done by five employees. Jennifer Gonzales performs Electron Microscopy for the team. Electron Microscopy (EM) may be used on a specimen when identification in cell culture is inconclusive to confirm the virus type. A submitter may also send clinical specimens for EM with suspect viral etiology or when other confirmatory tests are incomplete for pathogen identification.

Specimen grid preparation for viewing under the electron microscope. Photo courtesy of Andrew Vinyard.
Jennifer at the electron microscope. Photo courtesy of Andrew Vinyard.

“The 2013-14 influenza season kicked off in October, started off slow and steady, increased rapidly in December and is still going strong,” said Van Cleave. So far this season, the DSHS Laboratory VI team has received more than 500 specimens. The predominant positive strain thus far is Influenza 2009 H1N1, and there are a small number of specimens positive for Influenza A (H3N2) and Influenza B. This mirrors the pattern observed at the national level.

The path a specimen takes through VI is dependent on the virus suspected and the type of testing needed (i.e., whether or not it will receive molecular testing or cell culture). The presence of Influenza, including identification of commonly circulating types/subtypes and novel strains, is detected using a laboratory technique called Real-Time Reverse Transcription Polymerase Chain Reaction (Real-Time RT-PCR). “When we receive an influenza specimen, we extract the RNA from the clinical specimen and perform Real-Time RT-PCR using the Centers for Disease Control and Prevention (CDC) Human Influenza Virus Real-Time RT-PCR Diagnostic Panel assay,” said Van Cleave. “This is a rapid molecular method for detecting a specific virus. We convert the RNA to DNA, amplify the DNA, and use specific primer/probe sets to target the virus.”

A laboratorian performs automated RNA Extraction. Photo courtesy of Andrew Vinyard.

In addition to performing Real-Time RT-PCR, a subset of the influenza positive specimens identified by PCR are selected and placed onto cell culture to grow an isolate. These isolates are sent to the CDC along with the original clinical specimen for further characterization. The CDC uses these results to compare circulating strains to vaccine strains, guide decisions regarding treatment, formulate a vaccine for the coming year, and look for antiviral resistance and novel influenza viruses.

One of the positive specimens the VI team sent to the CDC during the 2012-13 Influenza season was selected to be included as a component in the 2013-14 vaccine. The entire process from detection to vaccine takes most of a year.

In April 2012, a nasopharyngeal (NP) wash was collected from a patient with influenza-like illness and sent to a laboratory in Fort Worth. It was then forwarded to the DSHS Laboratory in Austin. VI isolated an Influenza virus in cell culture from the NP wash. Both the virus isolate and the NP wash were sent to the CDC for characterization via the California State Public Health Laboratory. (CDC contracts with other state public health laboratories to first grow the viruses before characterization at CDC in Atlanta.) The original clinical specimen (NP wash) was inoculated into eggs at the CDC, resulting in an egg isolate identified as A/Texas/50/2012. Vaccine candidate virus was derived from the egg isolate and characterized at the CDC. An A/Texas/50/2012-like virus was recommended by WHO as the A(H3N2) component of Influenza vaccine in February 2013. The vaccine strain was sent to manufacturers for vaccine production.

Influenza surveillance specimens go through a final test in VI. “Currently, all of our positive and negative Influenza specimens are tested using the Luminex xTAG Respiratory Virus Panel (RVP) Assay, which is a multiplex test that can detect up to twelve different viral targets in a single well,” said Van Cleave “This assay is performed on positive Influenza specimens to determine if other viruses are present in addition to influenza (co-infection) and, in the case of negative influenza specimens, to determine if another virus is present that may have caused influenza-like illness.”

A laboratorian observing cell culture tube for cytopathic effect (CPE). Photo courtesy of Andrew Vinyard.Close up of cell culture tube. Photo courtesy of Andrew Vinyard.

Influenza isn’t the only headline-generating virus. In summer 2013, a Measles outbreak resulted in 21 cases in North Texas. Specimens were sent to VI. Some specimens were set up on cell culture. In addition, the DSHS Laboratory also sent specimens to the Minnesota Department of Health Laboratory (MN DOHL) for Measles PCR testing. The MN DOHL has been contracted by the CDC/APHL to provide vaccine preventable disease testing services, including Measles PCR testing.

This highly contagious disease is caused by the measles virus and can spread rapidly in an unvaccinated population. Even though measles is considered eliminated in the United States and all of the Americas, it is still common in other countries.

The infectious period for Measles is long; it can be transmitted four days before and four days after rash onset. Symptoms begin seven to 14 days after a person is infected and can include fever, cough, runny nose, conjunctivitis, and coryza (the symptoms of a cold). Three to five days after symptoms start, a maculopapular (a flat, red area covered by small bumps) rash appears that starts on the face and spreads downward, and a fever may spike.

According to the CDC website, measles is so contagious that any child who is exposed to it and is not immune will probably get the disease. That is what happened this summer (2013). The source was eventually identified as a child who had recently returned from Asia. A large proportion of the cases associated with this outbreak occurred in unvaccinated individuals. “This is why it is so important to receive the MMR (Measles, Mumps, Rubella) vaccine,” said Van Cleave.

Containment is a time-intensive undertaking. After identification in VI, epidemiologists in the DSHS Infectious Disease Section responded to measles cases by identifying all contacts within the nine-day infectious period, alerting them about their exposure, assessing the vaccination status of all the contacts, offering vaccine to those who were unvaccinated, and quarantining unvaccinated individuals for up to three weeks.

The work of the VI team is essential to containing viral outbreaks. The communicable nature of these viruses requires rapid turnaround of testing and results to appropriate infectious disease personnel to prevent further spread of disease.

by Jimi Ripley-Black


Newborn Screening Tour Aids India

Photo of tour attendees and guides. Photo courtesy of Andrew Vinyard.

Six visitors from India toured the DSHS Newborn Screening (NBS) Laboratory on December 4, 2013. These special visitors, who had been instrumental in starting a newborn screening program in India, were here to observe and learn from the testing operations of one of the largest newborn screening programs in the United States.

While at the Laboratory, tour attendees were provided with a NBS and clinical care overview, and then they visited NBS check-in, NBS testing areas, Laboratory Reporting and Container Preparation. The tour also included information about the Texas NBS screening laboratory infrastructure, quality assurance programs, staffing requirements and both health care provider and parent public education in regards to newborn screening.

Photo of tour from India in progress. Photo courtesy of Andrew Vinyard.

After the initial tour, one of the attendees, Dr. Gurjit Kaur, returned the following week for more in-depth observations of the NBS testing areas and training in hemoglobinopathy screening.




by Lucindra "Cindy" Corrigan



Celebrating 10 Years in the Laboratory Building

Laboratory managers hold the left side of the ribbon for the ribbon-cutting ceremony during the Grand Opening in 2003. Photo courtesy of Ivan Dudik.Employees hold over-sized scissors for the ribbon-cutting ceremony at the Laboratory Building Grand Opening in 2003. Photo courtesy of Ivan Dudik.Laboratory managers hold the right side of the ribbon for the ribbon-cutting ceremony during the Grand Opening in 2003. Photo courtesy of Ivan Dudik.

On November 1, 2013, laboratorians gathered to celebrate the 10th anniversary of the dedication of the new Laboratory building. Laboratory Display Boards were featured, illustrating each team's work and purpose and/or accomplishments.

Staff who were present during the relocation reminisced with visiting retirees over cake and punch. Perhaps the building isn't so new anymore, but for most long-term employees the move is a vivid memory that could have happened just yesterday. The transition to our new accommodations didn't happen all at once; it was accomplished in stages as areas of the Laboratory building were completed.

The first group to move over from the old building was Media Preparation, starting October 21, 2002, followed closely by personnel and business services, cost accounting, and procurement services. Serology, Newborn DNA Analysis and Prenatal Screening, and Prenatal Testing also completed their moves by the end of October. Specimen Acquisition was the first to move to the new building in November. For Chemist Roza Darghai, the Clinical Chemistry move date was memorable for more than one reason: "It was back in November 2002! That’s my starting date here at DSHS, and I remember we moved to the new lab only two days after I started my employment." Container Preparation moved at the end of November.

Serving cake at the 10th Anniversary Celebration of the Laboratory Building Grand Opening. Photo courtesy of Ivan Dudik.

Relocations continued in January 2003 as Newborn Screening and Lead Screening moved to the new building. For the Arbovirus team, the move was a bit too eventful. "It was spring, because I remember it was real windy and when we were moving the empty mouse cages they ended up being blown off our carts and all over the place," said Joanne Day. The rest of the Microbiological Sciences Branch, including Virology, trickled over to the new building by teams. Virology finally got to move in June.

The seventh floor of the Laboratory building wasn't finished for a couple of years after the Grand Opening celebration, so the final group, Environmental and Organic Chemistry, moved to the building between June and August 2005.

by Jimi Ripley-Black







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January 2014, Volume Five, Issue Three                          (Publication #E14-13156)
Published by DSHS Laboratory Services Section
PO Box 149347, MC 1947
Austin, TX 78714

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Grace Kubin, PhD
512 458 7318
email Grace

Jimi Ripley-Black
512 458 7318, ext 6505
email Jimi

Last updated April 28, 2014