Tuberculosis laboratory findings: Difference between revisions

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*Diagnostic Microbiology ([[pleural fluid]]): A sample of [[pleural]] [[exudate]] can be analyzed by [[cytopathology]] or at a cell count lab.  Samples are usually [[lymphocyte]] predominant, and cytopathology is more accurate than cell count labs at detecting lymphs.  If there is more fluid present, then an AFB lab is more appropriate.  A pleural exudate lab test may find sterile [[pyuria]] (especially in [[HIV]] positive patients), but overall this finding is fairly uncommon.
*Diagnostic Microbiology ([[pleural fluid]]): A sample of [[pleural]] [[exudate]] can be analyzed by [[cytopathology]] or at a cell count lab.  Samples are usually [[lymphocyte]] predominant, and cytopathology is more accurate than cell count labs at detecting lymphs.  If there is more fluid present, then an AFB lab is more appropriate.  A pleural exudate lab test may find sterile [[pyuria]] (especially in [[HIV]] positive patients), but overall this finding is fairly uncommon.
:Note: Most extra-pulmonary TB is pauci-bacillary, so the yield of tests is very low. This means that negative cultures do not mean no disease (e.g. negative [[cerebrospinal fluid]] AFB or even MTD is not that sensitive).
:Note: Most extra-pulmonary TB is pauci-bacillary, so the yield of tests is very low. This means that negative cultures do not mean no disease (e.g. negative [[cerebrospinal fluid]] AFB or even MTD is not that sensitive).
===Heaf Test===
The Heaf test is a [[diagnosis|diagnostic]] skin test performed in order to determine whether or not a child has been exposed to [[tuberculosis]]. Patients who exhibit a negative reaction to the test may be offered [[Bacillus Calmette-Guérin|BCG]] vaccination. The test is named after [[F. R. G. Heaf]].
Until 2005, the test was used in the United Kingdom to determine if the [[Bacillus Calmette-Guérin|BCG]] vaccine was needed; the [[Mantoux test]] is now used instead.  The Heaf test was preferred in the UK, because it was felt that the Heaf test was easier to interpret, with less inter-observer variability, and that less training was required to administer and to read the test.  The test was withdrawn because manufacturers could not be found for tuberculin or for Heaf guns.
The Heaf test may be informally referred to as the six pricks, as it gives six individual injections.
====Procedure====
A Heaf gun is used to inject multiple samples of testing serum under the skin at once. A Heaf gun with disposable single-use heads is recommended. 
The gun injects [[Mantoux test|purified protein derivative]] equivalent to 100,000 units per mL to the skin over the flexor surface of the left forearm in a circular pattern of six. The test is read between 2 and 7 days later. The injection must not be into sites containing superficial veins.
The reading of the Heaf test is defined by a scale:
*Negative - No induration, maybe 6 minute puncture scars
*Grade 1 - 4-6 papules (also considered negative)
*Grade 2 - Confluent papules form indurated ring (positive)
*Grade 3 - Central filling to form disc (positive)
*Grade 4 - Disc >10 mm with or without blistering (strongly positive)
Grades 1 and 2 may be the result of previous BCG or avian tuberculosis.
Children who have a grade 3 or 4 reaction require X-ray and follow-up.
The equivalent [[Mantoux test]] positive levels done with 10 TU (0.1 mL 100 TU/mL, 1:1000) are:
*0-4 mm induration (Heaf 0-1)
*5-14 mm induration (Heaf 2)
*>15 mm induration (Heaf 3-4)
The [[Mantoux test]] is preferred in the United States for the diagnosis of tuberculosis; multiple puncture tests, such as the Heaf test and [[Tine test]], are not recommended.


===Drug Resistance===
===Drug Resistance===

Revision as of 17:34, 4 September 2014

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Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1]; Associate Editor(s)-in-Chief: Alejandro Lemor, M.D. [2]

Overview

Laboratory Findings

Microbiological Studies

A definitive diagnosis of tuberculosis can only be made by culturing Mycobacterium tuberculosis organisms from a specimen taken from the patient (most often sputum, but may also include pus, CSF, biopsied tissue, etc). A diagnosis made other than by culture may only be classified as probable or presumed.

Distinctive clusters of colorless Mycobacterium tuberculosis form in this culture.

Sputum smears and cultures should be done for acid-fast bacilli if the patient is producing sputum. The preferred method for this is fluorescence microscopy (auramine-rhodamine staining), which is more sensitive than conventional Ziehl-Neelsen staining.[1]

If no sputum is being produced, specimens can be obtained by inducing sputum, gastric washings, a laryngeal swab, bronchoscopy with bronchoalveolar lavage, or fine needle aspiration of a collection. A comparative study found that inducing three sputum samples is more sensitive than three gastric washings.[2]

Other mycobacteria are also acid-fast. If the smear is positive, PCR or gene probe tests can distinguish M. tuberculosis from other mycobacteria. Even if sputum smear is negative, tuberculosis must be considered and is only excluded after negative cultures.

Many types of cultures are available [3]. Traditionally, cultures have used the Löwenstein-Jensen (LJ), Kirchner, or Middlebrook media (7H9, 7H10, and 7H11). A culture of the AFB can distinguish the various forms of mycobacteria, although results from this may take four to eight weeks for a conclusive answer. New automated systems that are faster include the MB/BacT, BACTEC 9000, and the Mycobacterial Growth Indicator Tube (MGIT). The MODS culture may be a faster and more accurate method [4].

  • Diagnostic Microbiology (sputum): The presence of acid-fast-bacilli (AFB) on a sputum smear or other specimen often indicates TB disease (at least 10,000c is needed on the smear to get a postive acid fast bacilli (AFB) stain). Acid-fast microscopy is easy and quick, but it does not confirm a diagnosis of TB because some acid-fast-bacilli are not M. tuberculosis. Therefore, a culture is done on all initial samples to confirm the diagnosis. However, a positive culture is not always necessary to begin or continue treatment for TB because cultures can take up to 3 weeks to yield definite results. A positive culture for M. tuberculosis confirms the diagnosis of TB disease. Culture examinations should be completed on all specimens, regardless of AFB smear results. Laboratories should report positive results on smears and cultures within 24 hours by telephone or fax to the primary health care provider and to the state or local TB control program, as required by law. A mycobacterium tuberculosis direct test (MTD) of nucleic acid amplification can also be performed to diagnose TB. An MTD test is similar to a polymerase chain reaction (pcr) and is very specific. The test is more sensitive than a smear but it is less senstive than a culture, and has the benefit of same day results.
  • Diagnostic Microbiology (pleural fluid): A sample of pleural exudate can be analyzed by cytopathology or at a cell count lab. Samples are usually lymphocyte predominant, and cytopathology is more accurate than cell count labs at detecting lymphs. If there is more fluid present, then an AFB lab is more appropriate. A pleural exudate lab test may find sterile pyuria (especially in HIV positive patients), but overall this finding is fairly uncommon.
Note: Most extra-pulmonary TB is pauci-bacillary, so the yield of tests is very low. This means that negative cultures do not mean no disease (e.g. negative cerebrospinal fluid AFB or even MTD is not that sensitive).

Drug Resistance

For all patients, the initial M. tuberculosis isolate should be tested for drug resistance. It is crucial to identify drug resistance as early as possible to ensure effective treatment. Drug susceptibility patterns should be repeated for patients who do not respond adequately to treatment or who have positive culture results despite 3 months of therapy. Susceptibility results from laboratories should be promptly reported to the primary health care provider and the state or local TB control program.

References

  1. Steingart K, Henry M, Ng V; et al. (2006). "Fluorescence versus conventional sputum smear microscopy for tuberculosis: a systematic review". Lancet Infect Dis. 6 (9): 570&ndash, 81. doi:10.1016/S1473-3099(06)70578-3.
  2. Brown M, Varia H, Bassett P, Davidson RN, Wall R, Pasvol G (2007). "Prospective study of sputum induction, gastric washing, and bronchoalveolar lavage for the diagnosis of pulmonary tuberculosis in patients who are unable to expectorate". Clin Infect Dis. 44 (11): 1415–20. doi:10.1086/516782. PMID 17479935.
  3. Drobniewski F, Caws M, Gibson A, Young D (2003). "Modern laboratory diagnosis of tuberculosis". Lancet Infect Dis. 3 (3): 141–7. PMID 12614730.
  4. Moore D, Evans C, Gilman R, Caviedes L, Coronel J, Vivar A, Sanchez E, Piñedo Y, Saravia J, Salazar C, Oberhelman R, Hollm-Delgado M, LaChira D, Escombe A, Friedland J (2006). "Microscopic-observation drug-susceptibility assay for the diagnosis of TB". N Engl J Med. 355 (15): 1539–50. PMID 17035648.

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