7 patients developed systemic anaphylaxis and 2 angioedema (Ahlroth et al. 1995) [255]

Systemic anaphylaxis in 7 patients with angioedema/urticaria in 6, vomiting in 2, bronchial asthma in 1, and rhinoconjuntivitis in 1 (Blanco et al. 1994) [287].

Urticaria and lip pruritus or glossitis in 25 patients. Rhinitis in 3 patients , facial angioedema in 2 patients and anaphylaxis in 2 patients (Blanco et al. 1994) [288].

The prevalence of clinical reactions to avocado among 17 latex-allergic patients was: 72% with asymptomatic sensitivity; 16% with anaphylaxis ; 2% with pruritus and 4% with generalized urticaria (Blanco et al. 1999) [289].

The clinical data for 16 avocado-allergic patientes was: 6 patients had systemic anaphylactic episodes; 2 with urticaria; 4 with angioedema; 3 with bronchial asthma and 4 with oral allergy syndrome (Chen et al. 1998) [36].

Fresh avocado (Blanco et al 1994) [288] and (Blanco et al 1994) [287], (Lavaud et al 1995) [102], (Ahlroth et al. 1995) [255], (Telezdiaz et al. 1995) [962]

Dilution 1/10 of a commercially available avocado extract (Beezhold et al 1996) [281].

Avocado extracts prepared as follows. Avocado pulp was ground in a mortar with cold acetone and then defatted with the same solvent (3× 1:5 (wt/vo)] for 1 hour at 4° C). The dried residues were extracted with 0.1 mol/L sodium phosphate buffer (pH 7.0) in 0.15 mol/L NaCl (PBS) (1× 1:5 (wt/vol) for 1 hour at 4° C), and after centrifugation, the supernatants were dialyzed against H₂O and freeze-dried (crude extracts). To obtain protein preparations enriched in chitinases, the PBS extracts (supernatants) were salted-out with 80% saturated (NH₄)₂SO₄. The pellets were collected by centrifugation, dissolved in PBS buffer, and then the pH was brought to 3.5 by addition of 0.1 mol/L HCl. After centrifugation, the supernatants were dialyzed against distilled H₂O and freeze-dried. Purified class I (Prs a 1) and class II avocado chitinases. (Blanco et al 1999) [289]

Avocado extracts. The consumable parts of avocado fruits (400 g) were homogenized in 1 L acetone solution containing 10% TCA at -70 °C, gently stirred for one hour and centrifuged for 10 min (-20 °C, 36 000 g ). The pellet was resuspended in acetone at -70 °C and centrifuged again. Residual acetone was removed by lyophilization (Posch et al. 1999) [163]

Avocado extract and fresh fruit (Isola et al. 2003) [598]

The positive results of the tests were designated as follows: + ; positive surface reference (histamine control: 10 mg/ml), ++ ; positive reference, +++ > positive reference, and ++++ > twice reference (Lavaud et al 1995) [102]

Prick-prick SPT results were graded using the size of histamine (10 mg/ml) reaction as a reference as follows: 4+ (2x histamine wheal), 3+ (1x histamine wheal), 2+ (0.5x histamine wheal), 1+ (<0.5x histamine wheal), and 0 (negative). (Ahlroth et al. 1995) [255]

Histamine and the diluent were used as positive and negative controls respectively (Beezhold et al 1996) [281].

Patients with a negative SPT to extracts were tested by prick-to-prick with the fresh fruit (Isola et al. 2003) [598]

25 patients with latex allergy. 9 of them showed avocado hypersensitivity with positive SPT. (Blanco et al 1994) [288]

17 patients with immediate-hypersensitivity to avocado. All patients were SPT positive with avocado (cultivar Strong) and 14 with the Hass avocado variety. (Blanco et al 1994) [287]

17 patients with latex allergy, 10 of them with a clinical history of avocado allergy (Lavaud et al 1995) [102]

18 patients with latex allergy (Ahlroth et al. 1995) [255]

47 latex allergic patients. (Beezhold et al 1995) [281]

100 consecutive atopic patients with allergic rhinitis (Telezdiaz et al. 1995) [962]

18 latex-allergic patients with a clinical history of allergy to chestnut, avocado, or both (Blanco et al 1999) [289]

20 latex-allergic subjects (Posch et al. 1999) [163]

82 patients with latex allergy.(Isola et al. 2003) [598]

9/25 patients showed avocado hypersensitivity with a positive skin prick test. (Blanco et al. 1994) [288]

All patients had a positive SPT with avocado (cv. Strong) but only 14 had a positive SPT with cv. Hass. (Blanco et al. 1994)

10/17 patients with clinical history of avocado allergy had a positive SPT to avocado. (Lavaud et al 1995) [102]

7/11 patients had a positive SPT to avocado (Ahlroth 1995).

53% of the latex -allergic patients and only 7% of the control non allergic were SPT positive to avocado (Beezhold et al. 1995)

The avocado allergen Prs a 1 elicited positive SPT responses in 12 out of 18 (67%) latex- fruit allergic patients. By contrast, avocado class II chitinase did not show any SPT responses . (Blanco et al. 1999)

8 of the patients were SPT positive with avocado extract and 3 with a negative SPT to extract were positive by prick-prick to avocado (Isola et al. 2003)

12 subjects had a positive response (Posh et al. 1999)

Of the 100 atopic patients not selected for avocado sensitivity, 21 had positive SPTs to avocado. (Telezdiaz et al. 1995)

The pulp of avocado was extracted with NTE 1N solution: NaCl 1mol/L, Tris 0,1mol/lL ethylenediaminetetraacetic acid 0.01mol/L (Lavaud et al 1995) [102]

Avocado extracts (Posch et al. 1999)

RAST (Lavaud et al 1995) [102]

CAP System RAST FEIA. Levels of specific IgE higher than 0.35 kU/L (class 1 and up) were considered positive (Blanco et al. 1999)

17 patients with latex allergy (Lavaud et al 1995) [102]

18 latex-allergic patients allergic with a clinical history to chestnut, avocado, or both (Blanco et al. 1999)

20 latex-allergic subjects (Posch et al. 1999)

10/17 had RAST responses to avocado Patients were grouped as follows: Group 1 comprised serum samples from patients with a clinical history of latex allergy, no clinical evidence for a fruit allergy, positive prick test and RAST responses to latex, negative SPT responses to fruits; Group 2 comprised serum samples from 10 patients with a latex allergy and a fruit allergy (suggestive symptoms, positive prick test and RAST responses) (Lavaud et al 1995) [102]

3/18 were IgE negative (Blanco et al. 1999)

13 subjects had a positive response by CAP (Posch et al. 1999)

Proteins were separated on 15% acrylamide SDS-PAGE under reducing conditions (Lavaud et al 1995) [102]

The allergen extracts were separated by sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE) with a 5% stacking gel and a 13% polyacrylamide gel (Moller et al. 1998).

The proteins were transferred onto a nitrocellulose sheet. The nitrocellulose sheet was blocked with 3% (w/v) skimmed milk and incubated with patient sera (1:100). IgE antibody detection was performed with alkaline phosphatase-conjugated goat anti-human IgE (1:100) and blots were developed with AP Conjugate Substrate (Lavaud et al 1995) [102].

The proteins were transferred onto a nitrocellulose membrane (0.2 µm) using a semi-dry blotting apparatus. Membranes were cut and blocked with 5% (w/v) skimmed milk and 0.1% (v/v) Tween 20 in PBS. The strips were incubated with patient sera. After incubation with rabbit anti-human IgE (1:4000), biotinylated goat anti-rabbit IgG (1:6000) and streptavidin-horseradish peroxidase-conjugated (1:20000), the blots were stained with 3,3",5,5"-tetramethylbenzidine-dioctylsodiumsulphosuccinate (Moller et al. 1998).

All patients' sera from group 2 (fruit and latex allergy) reacted to avocado proteins. The most important allergen in the extract was at about 30 kDa. A minor band was found at 21 kDa in one serum. Two sera from group 1 (monoallergy to latex) reacted with the 30 kDa protein (Lavaud et al 1995) [102].

IgE binding components of 43, 52, 58 and 65 kDa were detected by 3, 5, 7 and 5 serea respectively. One serum showed IgE binding to a protein of 27 kDa (Moller et al. 1998).

Oral as well as gastro-intestinal symptoms after ingesting avocado fruit