Sensitivity to mugwort pollen allergens (Artemisia vulgaris)

The author has found that 42% of patients with pollinosis had positive skin reactions with mugwort (Artemisia vulgaris) pollen allergens. The majority of tested patients (139 out of 187) were also allergic to grass pollens. However, hypersensitivity to mugwort pollen allergens was isolated and did not accompany grass pollen allergy. The symptoms of pollinosis appeared in this group later than in patients sensitive to grass pollen allergens only (over 21 years of age in 71%). Bronchial asthma was diagnosed in 40% of these patients and allergic skin reactions in 25%. Sensitivity to mugwort pollen allergens was accompanied by the sensitivity to pollen allergens of Graminae family of plants in 80% of cases. The author suggests that sensitivity to mugwort pollen allergens is the second most frequent cause of the pollinosis and is diagnosed too rarely. Failures of desensitization in patients sensitive to pollen allergens of Graminae family of plants may often result from coexisting sensitivity to mugwort pollen allergens as this sensitivity produces not only season but perennial clinical symptoms in nearly 50% of patients. The author discusses also botanical relations and cross-reactions in allergy to mugwort and ragweed pollen allergens.     

Comparison between Urticaceae (Parietaria) pollen count and hay fever symptoms: assessment of a «threshold-value»

Summary A comparison betweenParietaria pollen count and allergic symptoms of rhino-conjunctivitis in the early season was used in utilized to determine a «threshold-value» for this pollen. Clinical data were obtained from diary-cards of 34 allergic patients and pollen data from a volumetric sampling, carried out by means of a Hirst-Burkard pollen-trap. A significant correlation (r=0.98) was found between pollen count and symptom scores. Mild symptoms were registered with concentrations above 10–15 pollens/m³. Severe symptoms occurred when pollen count exceeded 80/m³/24 h., and over 90% of patients recorded symptoms. The importance of the late reactions and of the total allergenic airborne content are emphasized.     

Pollinosis in trentino (Northern Italy). Aerobiological and clinical research

Summary A study to evaluate and define the atmospheric pollen concentration in Trentino was carried out through the aerobiologic sampling in three localities chosen according to their different climatic conditions.1375 patients with pollinosis living in Trentino were studied retrospectively over the period ranging from 1986 to 1988 and selected according to the area they came from.Results have proved that the most allergenic pollen types are the following: Poaceae, Urticaceae (Parietaria), Compositae (Artemisia) and the tree pollen of Betulaceae and Corylaceae (Alnus, Betula, Corylus), and that pollinosis caused by such pollen, types has different features and frequencies according to the different localities.As far as symptoms are concerned, our data shows that rhinoconjuntivitis is more frequent in those patients who are allergic toParietaria while asthma results being more frequent in patients who are allergic to tree pollen.     

Trend of herbaceous pollen diffusion and allergic sensitisation in Genoa, Italy

In order to investigate the relationship betweenallergenic load and allergic sensitisation prevalenceon a long period, we analysed the annual data ofherbaceous pollen airborne diffusion in Genoa(NW-Italy) for 17 years (1981–1997), with particularregard to the most important allergenic species:Poaceae, Parietaria, Artemisia, Ambrosia. Weevaluated the relative trend and compared it with theprevalence data of patients sensitisation to thesepollens in the same period.The analysis of annual pollen amount revealed asignificant rising trend (r = 0.73, p < 0.05) of Parietaria pollen (average 14.324 grains/m³/year),whereas Poaceae (2048/year) and Artemisia(170/year) pollen count did not show any significantchange. Ambrosia (average 70/year) has been rising since 1989 (r = 0.87, p < 0.05) and reached themaximum pollen count in the last three years.The prevalence of pollen sensitisation resulted to bevery high (about 75%) in our atopic patients withrespiratory symptoms and did not change significantlyin the years: Parietaria pollen allergy waspresent in about 70% of patients with at least oneSPT positive to pollens (and 55% of atopicsubjects), and this frequency, much higher than theone found in North Italy, showed low year-to-year variability.Among pollinotics patients, Poaceae cutaneoussensitisation was found at a quite stable rate in theabove mentioned period (about 51%), as well as thatto Artemisia pollen (8%). Ambrosia skinprick test positivity is very rare (about 0.5%),without any evident increase.The present study shows that, for some of theherbaceous species, we cannot demonstrate astraightforward relationship between the trend ofairborne pollen and the frequency of sensitisation inexposed people. However, in the case of Parietaria, the high level of pollen amount in theair is clearly related with the high rate of cutaneous positivity in patients, but it seems tosuggest a ``plateau' that is not modified by anyfurther increase in airborne pollen (perhaps forgenetic limiting factors). On the other hand, Ambrosia pollen count, even if increasing in Genoa'satmosphere, could still be too low, and it would takesome more time to cause allergic sensitisation in exposed people.This kind of study, pointing out the influence ofairborne allergen exposure on hypersensitivitymechanisms, should be extended in time and world-wideto obtain general conclusions.     

The partial contribution of specific airborne pollen to pollen induced allergy

The air that we inhale contains simultaneously a multiple array of allergenic pollen. It is well known that such allergens cause allergic reactions in some 15 of the population of the Western World. However little is known about the quantitative aspect of this phenomenon. What is the lowest concentration of pollen that might trigger allergic responses? As people are exposed to heterogeneous and variable environments, clarification of the partial contribution of each of the major airborne pollen allergens and determination of its role in invoking allergy are of prime importance. Objectives: (1) Assessment of a possible correlation between the concentration of airborne pollen and incidence of allergy. (2) Estimation of the lowest average concentrations for various species of airborne pollen that elicit allergic symptoms when exceeded. (3) Determination of the extent of the variations in manifestation of allergy symptoms that can be explained by fluctuations in the concentration of individual species of airborne pollen. Methods: The study was conducted during 14months with a rural population in Israel. The participants completed a detailed questionnaire and were skin prick tested with the common airborne allergens. The appearance of clinical symptoms, i.e. nasal, bronchial, ocular or dermal, were reported daily by the patients. Concentrations of the airborne pollen and spores were monitored in the center of activity of the residents during one day every week, using three Rotorod pollen traps. The pollen grains were identified by light microscopy. Results: The pollen spectrum was divided into time-blocks presenting the main pollination periods of the investigated species. The correlation between the concentration of airborne pollen of the relevant species and the clinical symptoms of the patients was determined for each time block. The correlation differed for different clinical symptoms and for different pollen allergens. Highest correlation with airborne pollen counts was found for patients with nasal and bronchial symptoms. The onset of the clinical symptoms by sensitive patients started, in each of the relevant groups, once the weekly average concentration of the airborne pollen crossed a threshold level. Under the limitations of the present study, this level was estimated to be 2–4 pollen m^–3 air for olive, 3–5 pollen m^–3 air for grasses, 4–5 pollen m^–3 air for Artemisia, 10–20 pollen m^–3 air for pecan and 50–60 pollen m^–3 air for cypress. Conclusions: Fluctuations in specific airborne pollen grains explained up to 2/3 of the variation in clinical allergy responses. Those were: 69 of the variation for cypress (March–April), 66 for the grasses (March–April), 49 for the pecan (May–June) and 62 for Artemisia (Autumn).     

Allergen Microarray Indicates Pooideae Sensitization in Brazilian Grass Pollen Allergic Patients

Background

Grass pollen, in particular from Lolium multiflorum is a major allergen source in temperate climate zones of Southern Brazil. The IgE sensitization profile of Brazilian grass pollen allergic patients to individual allergen molecules has not been analyzed yet.

Objective

To analyze the IgE sensitization profile of a Brazilian grass pollen allergic population using individual allergen molecules.

Methods

We analyzed sera from 78 grass pollen allergic patients for the presence of IgE antibodies specific for 103 purified micro-arrayed natural and recombinant allergens by chip technology. IgE-ELISA inhibition experiments with Lolium multiflorum, Phleum pratense extracts and a recombinant fusion protein consisting of Phl p 1, Phl p 2, Phl p 5 and Phl p 6 were performed to investigate cross-reactivities.

Results

Within the Brazilian grass pollen allergic patients, the most frequently recognized allergens were Phl p 1 (95%), Phl p 5 (82%), Phl p 2 (76%) followed by Phl p 4 (64%), Phl p 6 (45%), Phl p 11 (18%) and Phl p 12 (18%). Most patients were sensitized only to grass pollen allergens but not to allergens from other sources. A high degree of IgE cross-reactivity between Phleum pratense, Lolium multiflorum and the recombinant timothy grass fusion protein was found.

Conclusions

Component-resolved analysis of sera from Brazilian grass pollen allergic patients reveals an IgE recognition profile compatible with a typical Pooideae sensitization. The high degree of cross-reactivity between Phleum pratense and Lolium multiflorum allergens suggests that diagnosis and immunotherapy can be achieved with timothy grass pollen allergens in the studied population.     

Localisation of allergens in ryegrass pollen and in airborne micronic particles

Summary In Melbourne, Australia, grass pollen allergens, especially from ryegrass, are a major cause of allergic hayfever and asthma. This review outlines recent developments in our understanding of how grass pollen allergens find their way into the atmosphere and how they are transported in particulate form. Much of this work has relied on antibody technology in immunological and immunocytochemical investigations. The localisation of allergens in situ has proved difficult due to their water-soluble character. Recently, allergens have been localised in developing ryegrass pollen by dryfixation, rapid-freeze and freeze-substitution techniques. This involved anthers being substituted in a mixture of aldehydes, organic solvents, and 2,2-dimethoxypropane. Incubation in dimethylsulfoxide prior to embedding in LR Gold resin provided good infiltration with freeze-substituted material. Immunogold-labelled sections show that the major allergens, Lol p 1 and Lol p 5, are synthesised in the pollen cytoplasm from the early bicellular stage, soon after the first starch granules are formed. From the early tricellular stage, Lol p 5 moves into the starch granules where it remains until maturity. Lol p 1 is localised in the cytoplasm of mature pollen grains. The incidence of airborne grass pollen, as measured in pollen traps, correlates with hayfever symptoms. Forecasting models which rely on rainfall and temperature data have been produced for the grass pollen (daily and seasonal) counts in Melbourne. Research over the past six years has shed light on the causes of grass-pollen-induced asthma. Micronic particles in the atmosphere may be starch granules originating from pollen grains osmotically ruptured by rainwater. Ultrastructural and immunological characterisation of micronic particles collected from outdoor air filters confirm the presence of airborne starch granules. These are loaded with grass pollen allergens, occur in the atmosphere especially after rainfall, and correlate significantly with instances of allergic asthma. Diesel particles might also play a role in the transmission of grass pollen allergens and thus become an extra asthma trigger. A variation in the mode of release of micronic particles occurs in other species, such as birch, where such particles are derived from burst birch pollen tubes. These particles are positive for Bet v 1 and are starch granules which are released into the atmosphere after light rain as a result of pollen germination on, e.g., leaves. After subsequent rupture of pollen tubes their contents are released when conditions become drier.Abbreviations DECP diesel exhaust carbon particles - DMP 2,2-dimethoxypropane - GPC grass pollen count - IgE immunoglobulin E - IgG immunoglobulin G - OGPS onset of the grass pollen season     

Molecular characterization of polygalacturonases as grass pollen-specific marker allergens: expulsion from pollen via submicronic respirable particles

Grass pollen belong to the most important allergen sources involved in the elicitation of allergic asthma. We have isolated cDNAs coding for Bermuda grass (Cynodon dactylon) and timothy grass (Phleum pratense) pollen allergens, belonging to a family of pectin-degrading enzymes (i.e., polygalacturonases). The corresponding allergens, termed Cyn d 13 and Phl p 13, represent glycoproteins of approximately 42 kDa and isoelectric points of 7.5. rPhl p 13 was expressed in Escherichia coli and purified to homogeneity. Immunogold electron microscopy using rabbit anti-rPhl p 13 Abs demonstrated that in dry pollen group 13, allergens represent primarily intracellular proteins, whereas exposure of pollen to rainwater caused a massive release of cytoplasmic material containing submicronic particles of respirable size, which were coated with group 13 allergens. The latter may explain respiratory sensitization to group 13 allergens and represents a possible pathomechanism in the induction of asthma attacks after heavy rainfalls. rPhl p 13 was recognized by 36% of grass pollen allergic patients, showed IgE binding capacity comparable to natural Phl p 13, and induced specific and dose-dependent basophil histamine release. Epitope mapping studies localized major IgE epitopes to the C terminus of the molecule outside the highly conserved functional polygalacturonase domains. The latter result explains why rPhl p 13 contains grass pollen-specific IgE epitopes and may be used to diagnose genuine sensitization to grass pollen. Our finding that rabbit anti-rPhl p 13 Abs blocked patients' IgE binding to the allergen suggests that rPhl p 13 may be used for immunotherapy of sensitized patients.     

Bronchial asthma in hay fever

The author diagnosed cough, emphysema and the symptoms characteristic for the bronchial asthma in 43% of patients with seasonal allergy (rhinitis, conjunctivitis) to pollens. Such symptoms were more frequent (51% of cases) in patients allergic to the grass pollens with coexisting hypersensitivity to Compositae family. Asthmatic symptoms in patients allergic only to grass pollens were seen in 38%. The author suggests that prolonged exposition in the inhalatory allergens (from two to four-five months) plays an important role in asthma onset in such patients. It rather delays than accelerates admittance to allergic clinics.     

Oleaceae in Trieste (NE Italy): aerobiological and clinical data

Oleaceae pollen concentrations in the Trieste area are low (<10%) compared to the total number of airborne pollen grains, with only one pollination peak at the beginning of June when the Oleaceae concentration reached 62% of the total pollen count in 1992. This peak was due to an exceptionally high pollination of olive trees in 1992, when airborne pollen concentration rose to 1357 pollen grains/m³ on June 3. Sensitization toOlea increased from 1989 to 1993, when it was present in 23.4% of symptomatic patients, but the role ofOlea in inducing allergic respiratory symptoms is difficult to evaluate because almost all patients were sensitized to other pollens, and in particular to Gramineae: only four subjects were sensitized toOlea alone (1.4%). Despite the low Oleaceae pollen counts, this pollinosis is increasing, probably because of increased cultivation ofOlea in recent years. However, although skin prick test positivity is common in polysensitized patients, its role in inducing sensitization and symptoms is presently less important than that of other pollens.     

The relationships between the concentrations of airborne pollen and allergic symptoms in Trieste (Northern Italy) in 1989

Summary Concentrations of airborne pollens recorded in Trieste in 1989 are evaluated in relation to allergic complaints in 113 patients with skin prick tests positive to one pollen species. Analysis of the result enable the two most important allergens to be confirmed as: Poaceae in 70.7% of patients and Parietaria in 18.6%. Few people had monosensitation to Compositae (Artemisia), Corylaceae and Fagaceae.Sympotoms are related to the flowering period when pollen levels climbed to daily averages of 15–20 grains/m³ but they started later than airborne peak concentrations and finished later than pollen decline. Pollen concentration recording can be a useful way to predict the clinical manifestation in sensitive patients but other factors are involved in determining symptoms like subjective mucosal reactions, polysensitization, patients living and working environment.     

Nonanaphylactic synthetic peptides derived from B cell epitopes of the major grass pollen allergen, Phl p 1, for allergy vaccination.

Worldwide more than 200 million individuals are allergic to group 1 grass pollen allergens. We have used the major timothy grass pollen allergen Phl p 1, which cross-reacts with most grass-, corn-, and monocot-derived group 1 allergens to develop a generally applicable strategy for the production of hypoallergenic allergy vaccines. On the basis of the experimentally determined B cell epitopes of Phl p 1, we have synthesized five synthetic peptides. These peptides are derived from the major Phl p 1 IgE epitopes and were between 28-32 amino acids long. We demonstrate by nuclear magnetic resonance that the peptides exhibit no secondary and tertiary structure and accordingly failed to bind IgE antibodies from grass pollen allergic patients. The five peptides, as well as an equimolar mixture thereof, lacked allergenic activity as demonstrated by basophil histamine release and skin test experiments in grass pollen allergic patients. When used as immunogens in mice and rabbits, the peptides induced protective IgG antibodies, which recognized the complete Phl p 1 wild-type allergen and group 1 allergens from other grass species. Moreover, peptide-induced antibodies inhibited the binding of grass pollen allergic patients IgE antibodies to the wild-type allergen. We thus demonstrate that synthetic hypoallergenic peptides derived from B cell epitopes of major allergens represent safe vaccine candidates for the treatment of IgE- mediated allergies.     

Clinical significance of allergic sensitization toOlea europaea L. pollen in Naples area,Italy

Summary The aim of this study is the evaluation of clinical significance of the allergic sensitization toOlea europaea L. pollen allergens in the Naples area, because in the last 10 years an increasing frequency of IgE mediated sensitization to these allergens has been observed. 4124 patients of both sexes (2989 adults and 1153 children) consecutively examined in our Allergological Center from January 1, 1988 to December 30, 1989 have been considered in this study. The patients were selected on the basis of a history of rhinitis (R) and/or asthma (A) and were evaluated by clinical examination, SPT, RAST and specific or aspecific BPT. Among 2267 adults and 879 children sensitized to almost one aeroallergen we found respectively 306 (13.49%) and 75 (8.53%) subjects with an allergic sensitization toOlea europaea L. pollen. The frequency of clinical manifestation was respectively R+A (58.16%), R (39.54%), A (2.28%) in adults; R+A (61.33%), R (26.66%), A (12%) in children. A monosensitization toOlea europaea L. pollen was found in 1.33% of children and in 2.28% of adults examined. The association with other allergens especially with mites and/or other pollen was very common.     

Identification of an allergen related to Phl p 4, a major timothy grass pollen allergen,in pollens,vegetables, and fruits by immunogold electron microscopy

Group 4 grass pollen allergens represent 60 kDa glycoproteins recognized by 70% of patients sensitive to these pollens. An antiserum against purified Phl p 4 from timothy grass pollen was used to investigate various pollens, fruits, and vegetables for Phl p 4-related allergens by immunogold electron microscopy. In timothy grass, mugwort, and birch pollens, allergens were located in the wall, and in timothy grass and birch pollens additionally in the cytoplasm. In peanut, apple, celery root, and carrot root, only cytoplasmic areas were labeled. Group 4-related allergens thus occur in pollens of unrelated plants and in plant food and may therefore contribute to crossreactivities in patients allergic to various pollens and plant food.     

Allergenic pollen and pollinosis in Warsaw

Summary Our study of fifty two hay fever patients included twenty six solely allergic to grass pollen and twenty six exhibiting allergy to various pollen species, such as hazel, birch, oak, poplar, andArtemisia. Their total and specific IgE response was evalutated by the immunoenzymatic method, while clinical reactivity was assessed by recording nasal and bronchial symptom scores between mid-March and mid-July. Simultaneously pollen counts were made. Polysensitized patients showed significantly higher levels of both total and specific IgE, which testifies to the enhanced quantitative and qualitative IgE. Multisensitized patients reacted earlier than patients sensitized to grass pollen only, which confirms that non-grass plants flowering only in the spring cause the priming effect on the nasal and bronchial mucosa. The early symptoms may be attributable to tree pollen sensitivity or may refletct higher grass pollen IgE levels in the polysensitized group. Characteristically, nasal symptoms preceded bronchial symptoms of several weeks.On comparing nasal washing from the polysensitized patients to washing from patients with grass pollen, we found much cytological material with the predominance of eosinophils.     

Transgenic ryegrasses (Lolium spp.) with down-regulation of main pollen allergens

Ryegrass pollen (Lolium species) is a widespread source of air-borne allergens and is a major cause of hayfever and seasonal allergic asthma, which affect approximately 25% of the population in cool temperate climates. The main allergens of ryegrass pollen are the proteins Lol p 1 and Lol p 2. These proteins belong to two major classes of grass pollen allergens to which over 90% of pollen-allergic patients are sensitive. The functional role in planta of these pollen allergen proteins remains largely unknown. Here we describe the generation and analysis of transgenic plants with reduced levels of the main ryegrass pollen allergens, Lol p 1 and Lol p 2 in the most important worldwide cultivated ryegrass species, L. perenne and L. multiflorum. These transgenic plants will allow the study of the functional role in planta of these pollen proteins and the determination of potential for development of hypo-allergenic ryegrass cultivars.     

Google Trends reflect allergic rhinitis symptoms related to birch and grass pollen seasons

Google Trends (GT) describes the variation of the relevant interest of internet searches toward medical conditions and related symptoms. Allergic rhinitis symptom levels result from the intensity of exposure to aeroallergens in combination with relevant medication use. We analyze data from Germany to examine the relationship between hay fever-related Google search terms, symptom levels, medication use, and pollen count levels. For doing so, we also employ the new definitions on pollen season and peak pollen period start and end as proposed by the European Academy of Allergy and Clinical Immunology in a recently published position paper. We extract GT data for a number of search terms related to allergic rhinitis for Germany. We use total nasal symptom and mediation scores as reported by patients via a patient hay fever diary in the Berlin and Brandenburg areas in Germany for 3 years (2014–2016), accompanied by pollen data. Then a Pearson and Spearman correlation analysis is performed between symptom data and GT data. A graphical analysis is conducted, and the identification of pollen season and peak pollen periods is done based on the EAACI criteria. The analysis reveals that GT data are highly correlated with symptom levels and follow peak pollen period start–end, concerning grass and birch pollen-induced allergic rhinitis symptoms. GT data can be used as a proxy for the identification of the onset and variation of nasal symptom and medication score for allergic rhinitis sufferers.     

The impact of pollen load on quality of life: a questionnaire-based study in Lithuania

Populations worldwide have been increasingly affected by allergic rhinitis associated with pollen allergens. Alterations in the geographical distribution of pollinating flora resulting from climatic changes and increased production of pollens caused by higher atmospheric CO₂, have changed the environment in which humans reside. This study, carried out in Lithuania, aimed to demonstrate how people with pollen allergy are affected by symptoms associated with changing pollen loads and to evaluate the resultant changes in human quality of life. This study assessed pollen loads in Lithuania during 2013–2014. Skin prick tests were carried out on 665 individuals aiming to evaluate sensitivity to pollen allergens. Self-reports on symptoms and their impact on quality of life were collected from each study subject. There were 24× to 42× differences in inter-annual peaks of Betula pollen concentrations, while differences of plant pollen concentrations of the other genus/families varied from 1.5 to 92 % on peak days. Positive reactions to pollen allergens were reported in 54 % of the study subjects; 90 % of these subjects had been continuously suffering from moderate-to-severe symptoms of allergic rhinitis. The most frequently reported symptoms were sneezing (99 %), runny nose (98 %) and stuffy nose (97 %). The dominant group of subjects reported symptoms during the entire period of vegetation (7 months). This study demonstrated that people with pollen allergies have attributed symptom-associated worsening of their quality of life to the typical periods of unavoidable exposure and have made only minimal changes in their lifestyle during the season of pollen load.     

Gain of structure and IgE epitopes by eukaryotic expression of the major Timothy grass pollen allergen, Phl p 1

Approximately 400 million allergic patients are sensitized against group 1 grass pollen allergens, a family of highly cross-reactive allergens present in all grass species. We report the eukaryotic expression of the group 1 allergen from Timothy grass, Phl p 1, in baculovirus-infected insect cells. Domain elucidation by limited proteolysis and mass spectrometry of the purified recombinant glycoprotein indicates that the C-terminal 40% of Phl p 1, a major IgE-reactive segment, represents a stable domain. This domain also exhibits a significant sequence identity of 43% with the family of immunoglobulin domain-like group 2/3 grass pollen allergens. Circular dichroism analysis demonstrates that insect cell-expressed rPhl p 1 is a folded species with significant secondary structure. This material is well behaved and is adequate for the growth of crystals that diffract to 2.9 A resolution. The importance of conformational epitopes for IgE recognition of Phl p 1 is demonstrated by the superior IgE recognition of insect-cell expressed Phl p 1 compared to Escherichia coli-expressed Phl p 1. Moreover, insect cell-expressed Phl p 1 induces potent histamine release and leads to strong up-regulation of CD203c in basophils from grass pollen allergic patients. Deglycosylated Phl p 1 frequently exhibits higher IgE binding capacity than the recombinant glycoprotein suggesting that rather the intact protein structure than carbohydrate moieties themselves are important for IgE recognition of Phl p 1. This study emphasizes the important contribution of conformational epitopes for the IgE recognition of respiratory allergens and provides a paradigmatic tool for the structural analysis of the IgE allergen interaction.     

New biomolecular tools for aerobiological monitoring: Identification of major allergenic Poaceae species through fast real‐time PCR

Grasses (Poaceae) are very common plants, which are widespread in all environments and urban areas. Despite their economical importance, they can represent a problem to humans due to their abundant production of allergenic pollen. Detailed information about the pollen season for these species is needed in order to plan adequate therapies and to warn allergic people about the risks they take in certain areas at certain moments. Moreover, precise identification of the causative species and their allergens is necessary when the patient is treated with allergen‐specific immunotherapy. The intrafamily morphological similarity of grass pollen grains makes it impossible to distinguish which particular species is present in the atmosphere at a given moment. This study aimed at developing new biomolecular tools to analyze aerobiological samples and identifying major allergenic Poaceae taxa at subfamily or species level, exploiting fast real‐time PCR. Protocols were tested for DNA extraction from pollen sampled with volumetric and gravimetric methods. A fragment of the matK plastidial gene was amplified and sequenced in Poaceae species known to have high allergological impact. Species‐ and subfamily‐specific primer–probe systems were designed and tested in fast real‐time PCRs to evaluate the presence of these taxa in aerobiological pollen samples. Species‐specific systems were obtained for four of five studied species. A primer–probe set was also proposed for the detection of Pooideae (a grass subfamily that includes also major cereal grains) in aerobiological samples, as this subfamily includes species carrying both grass allergens from groups 1 and 5. These, among the 11 groups in which grass pollen allergens are classified, are considered responsible for the most frequent and severe symptoms.