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TB Diagnosis: Line Probe Assay/ Hains Test

Introduction:

LPA(Line Probe Assay) or Hains test/ Molecular Methods is used to diagnose MDR/XDR TB Rapidly. This test is endorsed by WHO and FIND(Foundation for innovative new diagnostics.

Line Probe Assay Steps: 

  1.  DNA is extracted from M. tuberculosis isolates or directly from clinical specimens.
  2.  Polymerase chain reaction (PCR) amplification of the resistance-determining region of the gene under question is performed using biotinylated primers.

Features of LPA or Hains Test:

1. Rapid genotypic test based on DNA strip technology

2. Rapid dterminationof genetic mutatins associated with spcific 1st line & 2nd line drugs.

3. Line Probe Assay (LPA) technology is suitable for use at national/central reference laboratories, or at laboratories where there is proven capacity to conduct molecular testing

Resistance Vs Gene Mutation:

  • Rifampicin : rPOB
  • Isoniazid : KatG & inhA gene
  • Fluroquinolones : gyra gene
  • Aminoglycosides : rrs gene
  • Cyclic peptides : rrs gene
  • Ethambutol : embB gene 

Conclusion:

We recommend Hain test to diagnose MDR-TB in highly suspected cases, and to wait for conventional culture results to confirm NTM(Non-tuberculous Mycobacteria). Hain test has benefit in negative direct smear

References: 

1. www.ncbi.nlm.nih.gov
2.  https://www.tbfacts.org/drug-susceptibility/
3. "Molecular Line-Probe Assay for the Detection of Resistance to Second Line Anti-TB Drugs”, WHO, www.who.int/tb/areas-of-work/laboratory/policy_statements

Comments

  1. Jhon mac30 July 2019 at 11:17

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    Gene assay

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