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 Liquid Based Cytology 

Cervical cancer represents an important health problem at world level. It is the second worst cancer for women, with about 500.000 new cases per year. Moreover, the global world picture suffers from heavy disparities from the geographical point of view (Alliance for Cervical Cancer Prevention, 2004).

The 80% of all the new cancer cases are registered in developing countries. In many developing countries, the uterine cervix is one of the most relevant sites for cancer, comprising about 25% of all female cancers.

This is obviously due to the different planning and implementation of the screening programmes. As regards to vaccination programmes, it is not reasonable to think about an effective implementation of such programmes in the developing countries before ten years or more.

International Classification of Diseases-10 codes: C53

International Classification of Diseases-10 codes: C53

Also in industrialised populations the disease is common. In eastern and central European populations, the annual age-adjusted incidence rates for invasive disease are 15/25 per 100.000 women. In the Nordic countries, the annual incidence was 15/30 per 100.000 women, and is recently decreased thanks to the start of large-scale mass screening programmes. While cervical screening cannot be 100 per cent effective, cervical screening programmes have been shown to reduce the incidence of cancer in a population of women. For example the percentage of preventable cancers increased thanks to the new screening methodologies mainly for women aging from 20 to 39 years (Sasieni, Adams and Cuzick, BJC 2003). Worldwide, cervical cancer is the fifth most deadly cancer in women. It affects about 1 per 123 women per year and kills about 9 per 100,000 per year. Also the bladder cancer in Europe is relevant in statistical terms mainly for the men. It is the fourth most frequent cancer among men, accounting for about 7% of the total cancers. There are an estimated 136,300 new cases each year. The annual incidence rate is 32/100,000 in men and 9/100,000 in women while the annual mortality rate is 9/100,000 (11 for men, 4 for women) (Ferlay et al.). In Italy, bladder cancer incidence increased in both sexes (Zanetti, 2004). The incidence of kidney cancer increasing year by year in Europe (Figure 1) and the worrying trend highlight one more time the need to develop more and more the screening methodologies to minimise the detection error and decrease the number of false negative results.

Figure 1 - European kidney cancer incidence rate (Ferlay et al.)

Figure 1 - European kidney cancer incidence rate (Ferlay et al.)

All these diseases have a large worldwide diffusion and obviously the related screening costs are always increasing thanks to the even more massive application of the preventive medical policies by the different national health services.
Screening programmes are thus of fundamental importance in such areas, and concern two main possibilities: cytological observations and HPV testing.

Currently, cytological examination is one of the most used technologies with high effectiveness in screening examinations. Cervical cytology was introduced by the prominent Greek doctor George Papanicolau into clinical practice in 1940. The Papanicolaou test (also called Pap smear, Pap test, cervical smear, or smear test) is a screening test used in gynaecology to detect premalignant and malignant (cancerous) processes in the ectocervix. Significant changes can be treated, thus preventing cervical cancer. The test was invented by and named after Georgios Papanikolaou and in 1945 received the endorsement of the America cancer society as an effective method for the prevention of cervical cancer. The good results showed by this clinical practice in preventing many different and high diffused cancer typologies fostered the main players in this sector to invest resources in order to improve methodologies and result. Since the mid-1990s, techniques based around placing the sample into a vial containing a liquid medium which preserves the cells have been increasingly used. So far the most innovative way of preparing samples for cytological examinations in the laboratory is the Liquid-Based Cytology (LBC). In this kind of procedures the organic liquid to be investigated is settled on the glass microscope slide only after a preparatory procedure instead of the standard smearing deposition, proper of the conventional method.

In the other hand, Human Papilloma Virus (HPV) is responsible for cervix cancer or gene mutations and consequent hereditary diseases. According to the Alliance for Cervical Cancer Prevention, the use of HPV DNA testing will give women the opportunity for an early screening and detection of the cause of the cervical lesions. It is worth recalling that among the more than 120 known HPV types, 37 are known to be transmitted through sexual contact, and infection with sexually transmitted HPVs is very common in adult populations worldwide. In this regard, for example, the American Social Health Association projections in 2006 were so pessimistic as to predict that about 75% of the reproductive population will have been infected with genital HPV infection in their lifetime.Of course, not all the HPV genotypes are equally hazardous. Most of them come and go without ever causing symptoms. 13 HPV genotypes are classified as high-risk types and lead to the development of cervical cancer. Genotype 16, for example, is thought to cause 60% of the cervical cancers around the world; 10% of this pathology is instead related to the genotype 18, the second more hazardous genotype of the virus. Others, such as types 6 and 11, can cause genital warts.

Despite the fact the future availability of vaccines constitutes a great opportunity, screening still plays a fundamental role in limiting the health damage related to HPV also to check the effectiveness of the developed vaccines.


Our Method

CYTOfast system represents, since 1999, an important alternative to conventional cytological smear and to all the other LBC systems present on the market.

CYTOfast system main character is the vial filled by CYTOfast solution: this universal preservative solution allows the preservation of physiological structure and morphology of any kind of cells for 24 months at room temperature.

Cellular material left in the vial, after the slide preparation, can directly be used for further investigations with molecular biology techniques (e.g. PCR, hybridization, etc…).

CYTOfast alternative idea is represented by a standardization phase in which a nephelometric reading determinates the cellular density of the samples.
In accordance with it, the system fixes the quantity to add onto the slide for every sample, in order to obtain numerically standardized slides, always containing the same number of cells distributed as a monolayer, in a spot of 17 mm of diameter, for a safer, faster, easier and representative screening (˜ 100.000 cells). 

Slides are realized by aspecific centrifugation of cellular material; no filtration means that cells are not “chosen” on their size, therefore CYTOfast slide is 100% representative of the sample collected, for the most complete diagnosis.
On the contrary, many LBC methodologies are focused on the exploitation of the filtering technology, which basically presents the common advantages of LBC but some disadvantages too. Due to filtration, smaller cells (generally useful for detecting certain types of infections and inflammations) are lost in the process and, furthermore, the result is a slide that, as the first impression, results as “artefact”. 

Further on, our system is of the most competitive and cost-efficient system of the LBC market, thanks to our attention in reducing consumables and also thanks to our flexibility in offering the most attractive solution to our customers.
In this way, CYTOfast system provides all the LBC advantages, offering a valid alternative to other LBC systems further on.


CYTOfast Methodology


For gynaecological specimens, cells are collected from the patient with Rovers Cervex – Brush Combi or other suitable devices.

Brush head is assembled on its handle, then it is rotated in the patient three times.

The head of the brush is detached and put in the vial containing CYTOfast solution, in order to collect 100% of the cellular material.

For non gynaecological specimens, cellular liquids are collected in the vial containing CYTOfast solution without respecting any stoichiometric proportion.



Specimens containing the brush, once before their first processing, can be mixed on the Multimixer shaker, in order to allows the complete cells releasing in the preservative solution.

Multimixer shaker, designed for being compact, solid and user friendly, allows to shake up to 24 vials at the same time.


This phase, specific for Cytofast system, is realized by anephelometer that determinates the cellular density of the samples, allowing to collect a specific number of cells from the sample (˜ 100.000 cells), in order to realize a standardized monolayer slide, in a spot of17 mm of diameter, onto the slide.





Slides are realized by aspecific centrifugation of cellular material, to get a slide 100% representative of the sample collected.
Only with a specifically designed basculating platform is possible to “softly” push cells onto the slide: our system uses an Hettich cytocentrifuge model, 6 positions.




Polilysined slides are assembled with a specific centrifuge support and a plastic chamber, in order to realize the spot of 17 mmof diameter on them.
Fixation of cells onto the slide is realized by centrifugation force firstly, helped by the use of fixative solution, named Stickeur, and polylisined slides (electrostatic bonding).


Draining, staining and screening

After centrifugation, the supernatant of the slide is discharged, and slides are left to drain on air.
CYTOfast system doesn’t foresee any particular staining procedure; standard staining protocol is suggested to be use to every laboratory.

Screening phase of CYTOfast standardized monolayer slides results as safer, faster, easier and fully representative, if compared with conventional smears screening.
The high quality of CYTOfast slides allows sensible reduction of inadequate, border line and false negative cases.





Although CYTOfast system is born as an alternative to conventional Pap smear, it is a fully versatile system for all cytological specimens.
CYTOfast solution is a universal preservative solution, proper to collect and preserve all kinds of cytological materials.
The method is adapted for all the different cytological specimens, as Pap smear, urine, FNA sample, sputum etc.