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CBI - UOE > Invasive Pulmonary Aspergillosis (Bench to bedside diagnostics) > Flashcards

Invasive Pulmonary Aspergillosis (Bench to bedside diagnostics) Flashcards

1
Q

Aspergillus fumigateurs is the

A

most important opportunistic mould pathogen in immune-compromised humans

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2
Q

where are aspergillum fumigatus infections most common

A

haematological malignancy and allogenic bone marrow transplant patients

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3
Q

chronic pulmonary aspergilosis

A

3 million cases worldwide in patients with underlying lung diseases including asthma

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4
Q

allergic bronchopulmonary aspergillosis

A

4 million cases worldwide in patients with asthma and cystic fibrosis

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5
Q

everyone breathes in

A

spores- can get to lower part of respiratory tract

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6
Q

alveolar macrophages use which receptors to recognise fungal PAMP

A

PPR- Dectin-1

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7
Q

neutropenic bone marrow patients have

A

no immunity- zero WBC count

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8
Q

absence of macrophage and neutrophils in the lungs

A

dangerous

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9
Q

aspergilloma

A

a fungal ball- is a clump of mold which exists in a body cavity such as a paranasal sinus or an organ such as the lung. By definition, it is caused by fungi of the genus Aspergillus.

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10
Q

who is aspergilloma common in

A

those who have had TB and have scar tissue

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11
Q

symptoms of aspergilloma

A

weeping, snuggle to catch breath, cough up

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12
Q

CT scan of aspergilloma will show

A

mass within tissue

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13
Q

important to be able to differentiate between

A

normal breathed in spores and those that growing hyphae

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14
Q

process of pulmonary aspergillosis

A

1) inactive conidia are inhaled
2) conidia lodge in lower respiratory tract
3) condida swell
(block by macrophages
4) condida germinate into hyphae
(block by neutrophils)
5) hyphae invade tissue
(block by neutrophils)
6) hyphae invade blood vessels an disseminate

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15
Q

gold standard for IPA

A

no ‘gold standard’

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16
Q

imperative that diagnosis is

A

made without delay- prognosis worsens signify in the absence of recognition and effective treatment

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17
Q

CT scan and invasive pulmonary aspergillosis

A

cant see- hard to diagnpose

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18
Q

when someone comes to the doctor with IPA

A
  • their fever will be going up and down
  • fist thing doctor will do is give antibiotic
  • fever won’t resolve
  • then suspect fungal infection
  • window gets small for treatment
19
Q

treatment for IPA

A

horrendous side effects:
-hallucinations
-fever
due to fungal being very closely related to humans

20
Q

serological detection of aspergllus fumigatus

A

-elevated tigers of antibodies against A.fumigatus surface component

21
Q

elevated tigers in sera due to

A

presence of an abundant galactomannoprotein in the cell walls of the pathogen

22
Q

presence of an abundant galactomannoprotein in the cell walls of the pathogen

A

Elevated titers were shown to be due to the presence of an
abundant galactomannoprotein in the cell walls of the pathogen
(Afmp1p)

23
Q

AFMP1 gene cloned and sequenced and

A

recombinant Afmp1p

protein produced in E. coli

24
Q

what can be used to prove immunogneicity

A

Western blot

- purified Afmp1p protein antigen

25
Q

ELISA for the diagnosis of A.fumigatus aspergillosis

A
  • recombinant Afmp1p protein used to coat wells of micro titre plates
  • ELISA performed using sera taken from patients with aspergilloman, from patients with invasive aspergillosis and from patients with infections caused by the pathogenic fun
  • ELISA is highly specific for A.fumigatus diangosis
26
Q

when does ELISA not work so well

A

when patients are neutropenic- poor antibody response

27
Q

structures unique to fungi which are recognised by immune system

A
  • mannans and galactans- highly immunogenic

- making monoclonal antibodies against specific cell membrane proteins

28
Q

invasive pulmonary aspergillosis and galactomannans detection

A

double AB sandwich ELISA

29
Q

Invasive pulmonary aspergillosis and

galactomannan detection

A

Traditionally immunological tests for IPA have been centred
around the detection of the circulating fungal cell-wall
carbohydrate galactomannan (GM)
- using EB-A2 to detect

30
Q

reason for false positives within Invasive pulmonary aspergillosis and
galactomannan detection

A
  • does cross react with other fund e.g. Fusarium
  • cant tell if aspergillosis or invasive fusarium
  • baby milk contains galactomanna- moves into the gut and into the bloodstream
  • FM in penicillin
31
Q

Penicillin is very closely related to aspergillum

A

false positives- very dangerous

32
Q

cross reactivity of EB-A2 with G from other fungi

A

e.g. Fusarium

33
Q

Cross-reactivity of mAb with anti-cancer drug

A

cyclophosphamide

34
Q

‘Pan-fungal detection

A

when we do not know if a patient is infected with a virus, bacteria or fungi- don’t want to waste time treating for viral or bacterial

  • will pick up all fungi which affect humans
35
Q

Pan-fungal detection picks up

A

(1-3) B-D glucans from fungal cell wall

36
Q

(1-3) B-D glucans

A

Glucose polymers
Linked to proteins, lipids, mannan, chitin
Most fungi, some bacteria, most higher
plants, many lower plants
Up to 60% dry weight of fungal cells wall

37
Q

(1-3) B-D glucans from the basis of

A

fungi tell tests– high rate of false positives

38
Q

(1-3) B-D glucans does not detect

A

mucromycetes and cryptococcus that lack (1-3) B-D gluons in their cell walls

39
Q

Surrogate (non-GM) antigens for UPA detection

A

Alternative ‘circulating antigens’ are required as
surrogate markers for rapid diagnosis of IPA

Most appropriate targets are extracellular,
constitutively-expressed antigens

Should be able to discriminate between active
growth and quiescence

40
Q

which antibody is used

A

Mouse mAB JF5

- IgG3 immunoglobulin

41
Q

IgG3 immunoglobulin recognised

A

an extracellular constitutive, glycoprotein antigen

- antigen is secrets during active growth and not produced by dead spores

42
Q

mAbJF5- IgG3

A

displays superior specificity to rat mAb EB-A2

43
Q

which is the best marker for J5F (which is only produced when aspergillus fumigates is growing

A

Immunogold EM

CBI - UOE (61 decks)

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