Assay Sheet

  

Test ID

5000 Atypical Pneumonia Panel Qualitative PCR(includes Chlamydophila pneumoniae and Mycoplasma pneumoniae)

CPT Code

87486, 87581

Clinical Utility

Chlamydophila pneumoniae is a common cause of pneumonia throughout the world and causes 7% to 10% of community-acquired pneumonia among adults. Symptoms of infection with C. pneumoniae are indistinguishable from other causes of pneumonia. A physical examination or chest x-ray does not typically provide information which allows for a definite diagnosis.  Culture of the organism is technically demanding and time-consuming.  Alternatively, molecular assays such as PCR allow for rapid testing of respiratory secretions for the presence C. pneumoniae DNA, thereby allowing a diagnosis to be established and therapy instituted quickly.

Mycoplasma pneumoniae is the pathogen most often associated with atypical pneumonia. Often indistinguishable from other viral and atypical bacterial pathogens, M. pneumoniae causes a wide range of respiratory infections, including pneumonia, tracheobronchitis, and upper respiratory tract infection.  Specialized laboratory tests are necessary to establish a definitive diagnosis due to the nonspecificity of atypical pneumonia clinical presentation.  Traditionally, culture and serological tests have been utilized.  However, culture is relatively insensitive for acute diagnosis and requires a long incubation period.  PCR technology to assess for M. pneumoniae nucleic acid has proven to be a sensitive and specific diagnostic tool.  Respiratory specimens are tested for the presence M. pneumoniae DNA, facilitating a rapid diagnosis which allows appropriate antimicrobial therapy to be instituted quickly.

Procedure

Extraction of C. pneumoniae DNA and M. pneumoniae DNA from respiratory specimens followed by amplification and detection of C. pneumoniae DNA and M. pneumoniae DNA using real-time, qualitative PCR. An internal control is added to ensure the extraction was performed correctly and the PCR reaction was not inhibited.

Specimen type & specimen handling 

Bronchial Lavage/Bronchial Wash: 2 mls collected in a sterile, screw top tube. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.

Sputum: 2 mls collected in a sterile container, then transferred to sterile, screw top tube for shipment. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.

Throat Gargle:  2 mls collected in a sterile container then transferred to sterile, screw top tube for shipment. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.

Upper respiratory aspirate (NP aspirate, nasal aspirate/wash, tracheal aspirate, etc.): 2 mls collected in a sterile, screw top tube. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.

Upper respiratory swab (NP swab, throat swab): Sterile swab placed in 2 ml sterile saline, M4, or viral transport media in a sterile, screw top tube.  Do not use calcium alginate swab or wood shafted swab.  Ship at ambient temperature Monday thru Friday.  Specimen must be received within 96 hrs of collection.

All suction-type collection devices are inappropriate for specimen transport. Transfer specimen into sterile, leak proof tube for transport.

Call ViraCor for authorization prior to sending any specimen type other than those listed above.  

If another specimen type has received authorization for testing the following comment will appear in the final report: "The clinical utility of this result has not yet been demonstrated in the peer reviewed literature and is therefore unknown."

Causes for rejection

Wood shafted swab, calcium alginate swab.

Call ViraCor at 800-305-5198 if specimen is greater than 96 hrs old.

Specimen types other than those listed above that were sent without prior authorization.

Specificity

The Chlamydophila pneumoniae PCR assay was tested for cross reactivity against Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, all relevant non-C. pneumoniae species of Chlamydophila, all relevant strains of Mycoplasma pneumoniae, and all relevant species of Legionella as well as human herpes viruses, polyoma viruses, hepatitis viruses, adenoviruses, parvovirus B19, Pneumocystis jirovecii and Toxoplasma gondii with no cross reactivity noted.

The Mycoplasma pneumoniae PCR assay was tested for cross reactivity against Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, all relevant species of Chlamydophila, all relevant species of Legionella as well as human herpes viruses, polyoma viruses, hepatitis viruses, adenoviruses, parvovirus B19, Pneumocystis jirovecii and Toxoplasma gondii with no cross reactivity noted.

Assay Range

Qualitative results (Positive/Not Detected)

Turnaround Time

Same day (within 12 to 18 hours of receiving specimen), Monday through Saturday

Shipping

Ship Monday through Friday. Friday shipments must be labeled for Saturday delivery. All specimens must be labeled with patient's name and collection date. Multiple tests can be run on one specimen.

Ship specimens FedEx Priority Overnight® to:

ViraCor Laboratories, 1001 NW Technology Dr, Lee's Summit, MO 64086

The CPT codes provided are based on ViraCor's interpretation of the American Medical Association's Current Procedural Terminology (CPT) codes and are provided for informational purposes only. CPT coding is the sole responsibility of the billing party. Questions regarding coding should be addressed to your local Medicare carrier. ViraCor assumes no responsibility for billing errors due to reliance on the CPT codes illustrated in this material. PCR tests are performed pursuant to a license agreement with Roche Molecular Systems, Inc. This assay was developed and the performance characteristics were determined at ViraCor Laboratories. This test is performed in a CLIA certified laboratory. FDA approval is not required for the performance of this test.

1009 V1

Pathogen Overview

      

About Mycoplasma pneumoniae

Mycoplasma species are the smallest free-living organisms, with a genome size of 800kb, coding for 700 proteins.  These organisms are unique among prokaryotes in that they lack a cell wall, a feature largely responsible for their biologic properties such as their lack of a reaction to Gram stain and their lack of susceptibility to many commonly prescribed antimicrobial agents, including beta-lactams.  Mycoplasmal organisms are usually associated with mucosal surfaces, residing extracellularly in the respiratory and urogenital tracts. They rarely penetrate the submucosa, except in the case of immunosuppression or instrumentation, when they may invade the bloodstream and disseminate to different organs and tissues throughout the body. 

Clinical Manifestations

M. pneumoniae is the pathogen most often associated with atypical pneumonia. Often indistinguishable from other viral and atypical bacterial pathogens, M. pneumoniae causes a wide range of respiratory infections, including pneumonia, tracheobronchitis, and upper respiratory tract infection.  Only 3% to 10% of persons infected with M. pneumoniae develop pneumonia.  M. pneumoniae infections are more common in young children and older adults. Infection occurs throughout the year, but it can cause periodic outbreaks within small communities.  Transmission is by person-to-person contact, and infection spreads slowly, most often within closed populations such as hospital wards and intensive care units. Onset is insidious, over several days to one week. Symptoms include headache, cough, malaise, myalgias, sore throat, ear pain, and fever.  The cough is usually dry, paroxysmal, and worsens at night.  The clinical course of pneumonia caused by M. pneumoniae is usually mild and self-limited.  The mortality rate is approximately 1.4%.  However, pulmonary complications can be significant and involve respiratory distress.  Skin manifestations may include erythema and urticaria.  Neurologic derangements may include aseptic meningitis, cerebral ataxia, encephalitis, Guillain-Barré syndrome, and transverse myelitis.  The production of cold agglutinins can result in hemolytic anemia, especially when M. pneumoniae titers are high.  Additionally, complications such as myocarditis, pancreatitis, and polyarthritis can occur.

Diagnosis

Specialized laboratory tests are necessary to establish a definitive diagnosis due to the nonspecificity of atypical pneumonia clinical presentation.  Traditionally, culture and serological tests have been utilized.  However, culture is relatively insensitive for acute diagnosis and requires a long incubation period.  More recently, physicians have used serological testing for IgG and IgM to confirm M. pneumoniae infection.  However, definitive diagnosis with antibody tests requires seroconversion documented by paired specimens obtained 2 to 4 weeks apart.  More recently, PCR technology to assess for M. pneumoniae nucleic acid has proven to be a sensitive and specific diagnostic tool.  Patients with a strong clinical suspicion of mycoplasmal pneumonia should submit a respiratory sample for PCR testing of M. pneumoniae to facilitate early administration of appropriate antibiotics.

Treatment

Typically, treatment for pneumonia is initiated before the causative microorganism is identified. For empiric treatment, the most common type of antibiotic used is a macrolide such as azithromycin or clarithromycin.  Alternatively, fluoroquinolones may be administered.  Treatment is typically continued for 10 to 14 days for known infections. If the patient history involves symptoms for a prolonged period of time (> 6 months), the course of treatment should be a minimum of three weeks.

Selected References

Cunha BA. The atypical pneumonias: clinical diagnosis and importance. Clin Microbiol Infect. 2006;12(Suppl 3):12-24. 

Falguera M, Ruiz-Gonzalez A, Puig T, Nogues A, Garcia M. Detection of Mycoplasma pneumoniae by community-acquired pneumonia aspirates from patients with polymerase chain reaction in lung. Chest. 1996;(110):972-976.

Foy HM. Infections caused by Mycoplasma pneumoniae and possible carrier state in different populations of patients. Clin Infect Dis. 1993;17(Suppl 1):S37-46.

Klausner JD, Passaro D, Rosenberg J, et al. Enhanced control of an outbreak of Mycoplasma pneumoniae with azithromycin prophylaxis. J Infect Dis. 1998;177(1):161-166. 

Marston BJ, Plouffe JF, File TM Jr, et al. Incidence of community-acquired pneumonia requiring hospitalization. Results of a population-based active surveillance study in Ohio. The Community-Based Pneumonia Incidence Study Group. Arch Intern Med. 1997;157(15):1709-1718. 

Morozumi M, Ito A, Murayama S, Hasegawa K, Kobayashi R, Iwata S, Kawamura N, Kuroki H, Nakayama E, Tajima T, Ubukata K. Assessment of real-time PCR for diagnosis of Mycoplasma pneumoniae pneumonia in pediatric patients Canadian Journal of Microbiology. 2006; (52):125-129.

Nadal D, Bossart W, Zucol F, Steiner F, Berger C, Lips U, Altwegg M. Community-acquired pneumonia in children due to Mycoplasma pneumoniae: diagnostic performance of a seminested 16S rDNA-PCR. Diagnostic Microbiology and Infectious Disease. 2001;(39):15-19.

Nilsson A, Björkman P, Persson K. Polymerase chain reaction is superior to serology for the diagnosis of acute Mycoplasma pneumoniae infection and reveals a high rate of persistent infection. BioMed Central Microbiology. 2008;(8):93.

Parchuri S, Cunha BA. Mycoplasma pneumoniae community-acquired pneumonia: diagnostic usefulness of the agglutination-dissociation test. Infect Dis Pract. 2006;(30):550-551.

Pitcher D, Chalker V, Sheppard C, George R, Harrison T.  Real-time detection of Mycoplasma pneumoniae in respiratory samples with an internal processing control. Journal of Medical Microbiology. 2006;(55):149-155.

Suzuki S, Yamazaki T, Narita M, et al. Clinical evaluation of macrolide-resistant Mycoplasma pneumoniae. Antimicrob Agents Chemother. 2006;50(2):709-712. 

Talkington DF, Shott S, Fallon MT, Schwartz SB, Thacker WL. Analysis of eight commercial enzyme immunoassay tests for detection of antibodies to Mycoplasma pneumoniae in human serum. Clin Diagn Lab Immunol. 2004;11(5):862-867. 

Talkington DF, Waites KB, Schwartz S, Besser RE. Emerging from obscurity: Understanding pulmonary and extrapulmonary syndromes, pathogenesis, and epidemiology of human Mycoplasma pneumoniae infections. In: Scheld WM, Craig WA, Hughes JM, eds. Emerging infections. Vol 5. Washington, DC: ASM Press; 2001:57-84.

van Kuppeveld  F, van der Logt J, Angulo A, van Zoest J, Quint W, Niesters H, Galama J, Melchers W. Genus- and species-specific identification of mycoplasmas. Applied and Environmental Microbiology, 1992;58(8):2606-2615.

Waites KB. New concepts of Mycoplasma pneumoniae infections in children. Pediatr Pulmonol.  2003;36(4):267-278. 

Waites KB, Talkington DF. Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev. 2004;17(4):697-728.

Welti M, Jaton K, Altwegg M, Sahli R, Wenger A, Bille J. Development of a multiplex real-time quantitative PCR assay to detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae in respiratory tract secretions. Diagnostic Microbiology and Infectious Disease. 2003;(45):85-95.