Гематологические анализаторы крови широко используются для исследований крови при диагностике и мониторинге заболеваний. Большинство представленных на рынке медицинского лабораторного оборудования автоматических гематологических анализаторов позволяют проводить полный клинический анализ крови с расчетом лейкоцитарной формулы. Сложные и дорогие анализаторы могут оценивать клеточную морфологию и фиксировать клеточные популяции при диагностике редких заболеваний крови.
Принцип работы гематологических анализаторов крови
В основе устройства автоматических гематологических анализаторов лежат три технологии:
- Электрический импеданс
- Проточная цитометрия
- Флуоресцентная проточная цитометрия
Эти три метода предполагают использование химических реагентов, приводящих к лизису или изменения клеток для фиксации измеряемых параметров. Например, электрический импеданс позволяет дифференцировать эритроциты, лейкоциты и тромбоциты по объему. Добавление ядрообразующего агента, который сжимает лимфоциты больше, чем другие лейкоциты, позволяет дифференцировать объем лимфоцитов.
Электрический импеданс
Традиционным методом является метод электрического импеданса, иначе кондуктометрический или также называемый метод Коултера (Культера, Култера, Coulter) по имени его создателя. Этот метод/принцип используется практически во всех гематологических анализаторах.
Суть его состоит в том, что кровь пропускается между двумя электродами через отверстие настолько узкое, что через него может проходить только одна клетка. Импеданс или, проще говоря, проводимость среды изменяется по мере прохождения клеток через отверстие и это изменение пропорционально объему/размеру проходящих клеток. Данная зависимость и позволяет производить их дифференцированный подсчет.
Импедансный анализ позволяет выполнить клинический анализа крови с определением гранулоцитов, лимфоцитов и моноцитов, но он не позволяет различать гранулярные лейкоциты одинакового размера: эозинофилы, базофилы и нейтрофилы.
Скорость автоматического подсчета по данному методу в гематологических анализаторах составляет до 10000 клеток в секунду, и типичный анализ по импедансному методу может быть проведен менее чем за минуту.
Проточная цитометрия
Лазерная проточная цитометрия – более дорогостоящий метод по сравнению с импедансным, поскольку требует более дорогих реагентов, но он позволяет получать детальную картину морфологии клеток крови. Это лучший метод для определения лейкоцитарной пятикомпонентной формулы.
Суть метода состоит в том, что поток образца крови проходит через лазерный луч. Измеряется поглощение луча, а рассеянный свет измеряется под разными углами для определения зернистости, диаметра и внутренней сложности клетки. Это фактически те же самые морфологические характеристики клетки, которые можно определить вручную с помощью микроскопа.
Флуоресцентная проточная цитометрия
Добавление специальных флуоресцентных добавок позволяет расширить применение проточной цитометрии до возможности оценивать специфические популяции клеток. Флуоресцентные красители позволяют оценить соотношение ядро-плазма в каждой окрашенной клетке. Это используется для анализа тромбоцитов, зарождающихся эритроцитов и ретикулоцитов.
Обзор производителей гематологических анализаторов
Производители автоматических гематологических анализаторов крови объединяют в своих приборах эти три технологии с инновационным применением реагентов и методов обработки данных, создавая собственные технологии, каждая из которых имеет свои преимущества с точки зрения точности измерений, скорости их выполнения или набора параметров.
В приборах ADVIA ® фирмы Siemens используется образец пероксидазы для дифференцированного тестирования. С его помощью выполняется самотестирование прибора (внутренний QC тест).
Метод лаурилсульфата натрия, используемый в приборах фирмы Sysmex при определении гемоглобина является нецианидным методом и имеет очень короткое время реакции. Гемоглобин определяется в отдельном канале, при этом минимизируется влияние высоких концентраций лейкоцитов.
Анализатор CELL-DYN Sapphire ® от Abbott использует трехцветную флуоресценцию в сочетании с запатентованной технологией Multiangle Polarized Scatter Separation для обеспечения высочайшей точности при определении лейкоцитарной формулы за счет идентификации клеток с использованием четырех углов рассеяния света.
Что следует учитывать при покупке гематологического анализатора?
Выбор подходящего инструмента будет зависеть от того, где его предполагается использовать: возле постели пациента (прикроватный), в лаборатории для экспресс-диагностики, крупной клинической лаборатории или в исследовательском медицинском учреждении. Разумеется, нужно помнить, что оборудование, используемое в клинических целях, должно иметь соответствующие сертификаты.
Набор параметров, заложенных в автоматический анализатор
В каждый, даже самый простой автоматический гематологический анализатор крови заложена возможность определять не один, а несколько параметров крови. Все они выдают результаты клинического анализа CBC (complete blood count) и рассчитывают трех или пятикомпонентную лейкоцитарную формулу. Однако количество этих параметров у приборов разное. Наиболее простой гематологический анализатор XP-300™ от фирмы Sysmex дает результат по 17 параметрам: WBC; RBC; HGB; HCT; MCV; MCH; MCHC; PLT; NEUT #,%; LYM #,%; MXD #,%; RDW-SD; RDW-CV; MPV.
Модель же гематологического анализатора крови Pentra DX Nexus SPS от компании Horiba позволяет определять 50 параметров крови: WBC, RBC, HGB, HCT, MCV, MCH, MCHC, RDW, PLT, MPV, *PCT, *PDW %, NEU, LYM, MON, EOS, BAS, *ALY, *LIC, *IMM, *IML, *IMG: (% and #), RET (% and #), RETH %, RETM %, RETL %, *IMR %, CRC%, IRF, MRV, *RHCc, MFI, NRBC % NRBC#, *CWBC, *WBC#, *RBC#, *PolyNuc#, and *MonoNuc# и др. («*» отмечены те, что используются в исследовательских целях).
Делая выбор, необходимо представлять, насколько целесообразно приобретать прибор на большое количество параметров, поскольку и расходы на обслуживание и реагенты будут выше.
Время на анализ
Время анализа зависит от определяемого параметра и является одной из наиболее важных технических характеристик гематологического анализатора. Лимитирующей временной стадией является реакция образца с реагентом. Простой клинический анализ крови на автоматических гематологических анализаторах как правило выполняется за одну минуту, на определение более сложных параметров может уходить до 10 мин.
Анализатор CELL-DYN Sapphire от фирмы Abbot затрачивает на определение тромбоцитов иммунотромбоцитным методом CD 61 (immunoplatelet method) приблизительно 5 минут, а на определение CD3/4 и CD3/8 T-клеток около 7 минут.
Степень автоматизации
Высокопроизводительные лаборатории предъявляют высокие требования к степени автоматизации анализов. Рассматриваются такие характеристики анализатора, как максимальная загрузка образцами, скорость выполнения гематологического анализа, совместимость флаконов (виал или ячеек) с другим оборудованием.
Гематологический анализатор крови XE-5000 от Sysmex позволяет выполнять анализ образцов по 31 параметру со скоростью 150 образцов в час.
Гематологический анализатор крови ADVIA 120 от фирмы Siemens – настольный прибор, который позволяет загрузить в автосэмплер одновременно 150 образцов и выполнить их полный анализ без участия оператора за 75 минут.
Гематологический анализатор Pentra DX Nexus SPS от фирмы Horiba оснащён автозагрузчиком проб, совместимым с различными пробоподатчиками.
Сейчас необходимой технической характеристикой серьезного гематологического анализатора является возможность работать со штрих-кодами (бар-кодами). Технология считывания и распознавания данных у анализаторов разная.
Например у анализаторов крови LH 780 от Beckman Coulter реализована технология, которая позволяет достоверно считывать даже плохо пропечатанные изображения.
Особо следует отметить очевидную пользу от автоматической маркировки (выделения) результатов анализа, которые вышли за пределы допустимых диапазонов, возможность их автоматической перепроверки без участия оператора.
Расход и стоимость используемых реагентов в гематологических анализаторах
Заранее необходимо выяснить какие в гематологическом анализаторе применяются реагенты, в каком количестве и насколько они безопасны и экологичны. Важно узнать их основные характеристики, а также можно ли их приобрести у поставщиков реагентов или только у производителя.
Анализатор ELite 3 от компании Erba позволяет определять 20 параметров с использованием только трех реагентов, которые не содержат цианидов и вообще экологически безопасны.
В анализаторах DxH 800 и DxH 600 фирмы Beckman используется всего пять реагентов на определение всех параметров, включая NRBC и ретикулоциты.
Заранее выясняйте, как часто необходимо менять реагенты. Модель анализатора ADVIA 120 от Siemens поставляется с комплектом реагентов (анализ + промывка) на 1850 тестов.
Размер образца и работа с микроколичествами
Типичный размер аналитической пробы для проведения анализа составляет 150 μL. Некоторые производители позволяют выполнить анализ крови в пробе меньшего объема (микросэмпл), что очень востребовано в педиатрии. Например, компания Horiba выпускает анализатор модели ABX Micros ES 60, который может выполнить полный анализ крови используя всего 10 μL образца.
Точность, сходимость и линейность
Погрешность измерения объема клеток методом импеданса у большинства приборов может превышать 1% она определяется соотношением размера отверстия в импедансном узле к размеру исследуемых клеток. Некоторые приборы позволяют снизить эту погрешность за счет использования в измерительных модулях ячеек с разными диаметрами отверстий для клеток различного размера. Температура также влияет на погрешность. Особенность анализатора Pentra 80 от компании Horiba состоит в предварительном разогреве измерительного модуля, что повышает точность и стабильность результатов.
Время и периодичность технического обслуживания и калибровок
Узнайте, как часто анализатор необходимо обслуживать, калибровать и что входит в техническое обслуживание. Производитель Idexx в своих анализаторах задействует технологию qualiBeads®. Это частицы с определенными характеристиками, которые используются для проверки гематологического анализатора в качестве внутреннего стандарта.
В автоматических анализаторах крови ABX Pentra от Horiba заложена стандартизация, валидация, перепроверки, а также возможность обмена данными между центральной лаборатории и дочерними.
Образцы находятся в открытых или закрытых флаконах
Работа анализатора с закрытыми флаконами/виалами сводит к минимуму возможность пролива образца крови и тем самым возможную опасность от контакта оператора с пробой. Приборы, которые позволяют работать как с открытыми, так и закрытыми флаконами имеют различную стабильность результатов и механизмы калибровки в зависимости от выбранного режима.
Обработка результатов, их хранение
Количество результатов анализов, которые могут быть сохранены в системе у всех анализаторов разное. Компактные настольные анализаторы обычно позволяют сохранить до 1000 результатов и гистрогармм, а, к примеру, ABX Pentra DX120 SPS от Horiba позволяет записать 90000 результатов и графиков. Многие из высокопроизводительных систем интегрируются в LIMS.
Размеры гематологического анализатора
Размеры гематологических анализаторов варьируются, от компактных настольных моделей, до больших автоматизированных комплексов. Анализатор ABX Micros ES 60(Horiba) – компактная настольная модель с размерами 41 × 36 × 36 см. и весом всего 12,7 кг. Она управляется с тачскрина и позволяет записывать до 1000 результатов анализов.
Напольная высокопроизводительная модель XN-9000 от Sysmex собирается из модулей и конфигурируется под задачи заказчика. Конфигурация 801 имеет размеры 792 см × 122 см. Такой автоматизированный комплекс позволяет анализировать до 900 образцов в час.
Использование гематологических анализаторов в ветеринарии
Для автоматиечкого анализа крови животных используются специальные гематологические анализаторы. Анализатор ProCyte Dx от Idexx предназначены для анализа крови собак, кошек, лошадей, быков, хорьков, кроликов, песчанок, свиней, морских свинок и образцов мини-свиней.
Программное обеспечение Coulter Ac-T для ветеринарных программ от Beckman Coulter предназначено для обработки результатов анализов крови кошек, собак и лошадей.
Производители гематологических анализаторов
Самые широко известные производители это:
- Abbott Diagnostics
- Beckman Coulter
- Erba Diagnostics
- Horiba
- Idexx
- Siemens
- Sysmex
Менее популярные зарубежные производители:
- APPLIED BIOSYSTEMS (USA)
- BOULE MEDICAL AB (SWELAB) (SWEDEN)
- DIATRON MESSTECHNIK GMBH (AUSTRIA)
- DREW SCIENTIFIC, LTD (NETHERLANDS)
- F. HOFFMAN-LA-ROCHE LTD. (SWITZERLAND)
- HOSPITEX DIAGNOSTICS (ITALY)
- HUMAN GMBH (GERMANY)
Вот список российских компаний, которые производят автоматические гематологические анализаторы:
ООО "НПФ "Лабовэй" производит анализатор Пикоскель ПС-4М.
ООО «Эйлитон» производит гематологические анализаторы серии Medonic. Эта компания входит в группу компаний «А/О Юнимед» и работает на российском рынке медицинского лабораторного оборудования c 2003 года.
Анализ крови на них по 19 параметрам с производительностью 60 проб в час
НПФ ООО «ВИТАКО» производит анализаторы серии АРД.
Также на российском рынке представлены модели китайского производства.
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Статья и фотоматериалы взяты с сайта https://findlab.ru/
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