Autism is a common pervasive developmental disorder (PDD), the incident rate of which is about 2-5 cases / ten thousand children. According to the interviews with experts in relevant clinical institutions and public welfare organizations we have conducted, it is found that the public in mainland of China hold relatively poor awareness of this disease, causing delays in treatment of autistic children.Even in terms of the existing measures of testing and treatment, scale testing and behavioral intervention are dominant. However, this condition are in demand for development nowadays, for the reason that the accuracy of detecting and the strategy of treatment are likely to be polished as the technology develops. Especially, some researchers have found that probiotic beverage helps to assist the treatment of autistic children, which opens up oral medication therapy beyond behavioral intervention. Meanwhile, a study also has found that the contents of tryptophan, glutamate and copper ion in the blood of autistic patients increased significantly.

We hope that our project will shed light on the possibility and practicability of the diagnosis and treatment of autism at an early stage. In order to improve the contemporary situation, our team tries to develop a new method of diagnosing children who potentially have autism and curing them in a way of drinking probiotic beverages afterward.
Taking it as our aim, we have managed to do some basic experiments and reached the consequence we expected. In the future, our engineered probiotics will then be used for the construction of assay kits and therapeutic probiotic beverages for real-world application.

Based on further research, we find that SLC7A5 is a protein that can ease the autism symptoms. Therefore, we also constructed another plasmid, pET28a +SLC7A5 and then transform is into the competent bacteria E. coli BL21(DE3), also verified its expression level by SDS-PAGE and obtained the purified protein.

As seen in figure 3, we can see the Fluorescence intensity shows a positive correlation with the concentration of L-tryptophan in the pTrc99K-tnaC-amilGFP assay system which proves that this assay system could detect the changes in L-tryptophan so as to help detect the autism.

According to the SDS-PAGE results in figure 4, we could find our target proteins, inaK-gldh protein (left) and SLC7A5 protein (right), respectively, which also prove that our engineered plasmids both work as expected.

The one product of us is the healthy drink that added two probiotics we have cultivated. After taking the drink, the SLC7A5 protein will be secreted inside the body to assist in the treatment of autism. This kind of drink feature long-lasting effects, affordable price, and acceptability, with a good market prospect. Below is our design for this probiotic drink which could relieve the symptoms of autism.

The other product of ours is Autism Detection Kit. By measuring the level of glutamate and tryptophan, people could obtain the jointly diagnose result of autism. The kit, not like other current autism diagnosis methods, features the convenience of operation, accuracy of scientific detection, and low price. Below is our design for this autism detection kit.

Target customers of the products above mainly include the family with autistic children, institutions for autistic detection, and specialized hospitals for autistic children. As we have surveyed, there are almost no institutions and hospitals in China using such advanced or similar products for autism detection. Therefore, our products must be in great demand if our final products get into the market after proving their safety and effects.

Specifically, the target groups of our company's two products are similar, but there is a little difference. For the autism assay kit, the potential customers of this product are the people who have primary or suspected symptoms of autism but have not been diagnosed officially, and those who hope to check whether they are autistic. The probiotic drink aims to relieve autism symptoms, therefore this product is more targeting those people who have been identified as having autism.

The advancement of any science and technology should take safety as the basic guarantee, so safety must be guaranteed in the early, middle, and late stages of the entire process. Before the experiment, we must ensure that all experimental materials are safe, and they must be carefully checked and confirmed before they can be put into use. In the experiment, we not only need to ensure the security of the experimental environment, but also the specification of our operation steps. Finally, it is also very important that the intermediate and final products of the experiment are non-polluting to the environment and harmless to the human body.

Of course, risks and opportunities coexist, so recognizing the challenge is an important condition for us to solve it and even turn it into an opportunity. On one hand, since our final products are beverages and test boxes, we have to meet a series of government requirements for food and drug regulation. We have to go through enough clinical trials to prove its safety and effect on the human body, and we also need to obtain a special production license before formal industrial production. In addition, we also face some special requirements in promotion and sales. All require additional investigations and efforts.

On the other hand, it may face some challenges if distributed to the market since the probiotic drink needs to be drunk and the autism assay kit requires blood samples, which is likely to bring a psychological burden to parents. Considering this, we are prepared to conduct enough activities for science popularization aiming at developing the awareness and cognition of the public. In the meantime, we also need to spend more time promoting the safety of our products to eliminate the unknown concerns of our consumers for new products. In a word, the challenges and risks can be overcome through our active efforts.

For the assay kit, we plan to conduct more tests to obtain enough data for optimizing our models to predict a more exact concentration value of those mark elements by measuring the fluorescence intensity so as to increase its validity and accuracy. In addition, we shall do more literature investigation to settle down the reference value range of these mark elements for normal people so as to confirm the autism diagnosis reference.

For the probiotic drink, so far we have successfully expressed our engineered plasmid inside the E. coli. While in order to make the drink safer for the human body, we will switch to other host bacteria, like Nissle1917, which has been approved as the only edible non-pathogenic E. coli, to further test the performance.

Since the etiology of autism is a combination of congenital heredity, maternal-fetal development, and acquired environment, clinical symptom evaluation is mainly used to judge it. Our products undoubtedly provide a more convenient way to diagnose, but this diagnosis must also be combined with eye tracking, EEG detection and other methods to provide a reference for the diagnosis of autism. The task of developing new portable tests and effective treatments is imminent. In the future, when the biosensor is improved, it may be applied to ASD detection, which can provide a new tool for disease diagnosis, and the earlier the diagnosis is made, the better the outcome.

●ATL（Above the line）
For the promotion of our product, we will build websites and create official social media accounts to release product updates, answer questions regarding autism and our products, promote product utility, and popularize autism symptoms. Based on the huge number of social media users, we can let more people know the purpose of our products, attract potential users and establish trust with customers. By utilizing the platform, we can establish communication between patients and gain effective feedback from patients who use our products.

●BTL (Below the line )
Considering these difficulties that we might face when our company first enters the market, we can first build up our loyal customer group and our trust by promoting our products in hospitals. Besides, by holding performance activities and inviting autistic people to perform in talent shows, we can increase public awareness of autism patients, and increase our brand influence. In addition, taking the initiative to hold public welfare activities such as charity sales or free community medical services can help us to promote our products and our company. This method can also help some residents to detect whether they have autism and reflect the efficacy of our products.

●TTL(Though the line)
We also plan to promote the selling of some co-branded products with some snacks or toys favored by autistic patients and make some cartoons and videos to improve the popularity of the products and our brand reputation.

[1]Nadesan, Majia Holmer. “Autism and Genetics: Profit, Risk, and Bare Life.” Worlds of Autism: Across the Spectrum of Neurological Difference, edited by Joyce Davidson and Michael Orsini, University of Minnesota Press, 2013, pp. 117–42.
[2]Chen Yanyun, and Zhang Xiaolei. "Research progress of probiotics in the treatment of autism." China Medical Innovation 16.19 (2019): 169-172.
[3]Dora C. Tărlungeanu, et al. Impaired Amino Acid Transport at the Blood Brain Barrier Is a Cause of Autism Spectrum Disorder.cell.2016.